- Email: [email protected]
fork head winged helix family of transcription factors
GENOMICS
25,388-393
(19%)
Murine Chromosomal Location of Eight Members of the Hepatocyte Nuclear Factor 3/Fork Head Winged Helix Family of Transcription Factors KAREN
B. AVRAHAM,* COLIN FLETCHER,* DAVID G. OVERDIER,t DEREK E. CLEVIDENCE,t ROBERT H. CosTA,t NANCY A. JENKINS,* AND NEAL G. COPELAND*,’
ESENG LAI,+
*Mammalian Genetics Laboratory, ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Frederick, Maryland 2 1702; tDepartment of Biochemistry, College of Medicine, The University of Illinois, Chicago, Illinois 606 12; and *Division of Endocrinology and Cell Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, New York 1002 1 Received August 23, 1994; revised November 5, 1994
A 100.amino-acid DNA-binding motif, known as the winged helix, was first identified in the mammalian hepatocyte nuclear factor-3 (HNF-3) and Drosophila fork head family of transcription factors. Subsequently, more than 40 different genes that contain the winged helix motif have been identified. In the studies described here, we have determined the murine chromosomal location of eight members of this gene family, HFH-1, HFH-3, HFH-4, HFH-5, HFHS, HFH-8, BF-1, and BF-2, by interspecific backcross analysis. These genes, designated HNF-3 fork head homolog 1 (Hfhl), Hfh3, Hfh4, Hfh5, Hfh6, Hfh8, Hfh9, and HfhlO, respectively, mapped to 6 different mouse autosomes and are thus well dispersed throughout the mouse genome. Based on this mapping information, we predict the chromosomal location of these genes in humans and discuss the potential of these genes as candidates for uncloned 0 1996 Academic Press, Inc. mouse mutations.
INTRODUCTION
The study of cell type-specific transcriptional control has led to the identification of a class of transcription factors sharing a DNA-binding domain of the type found in hepatocyte nuclear factor 3 alpha (HNF-3a), HNF-3P, and HNF-3 y and in the Drosophila fork head (FKH) proteins. The nearly 40 genes identified so far (reviewed by Lai et al., 1993; Costa, 1994) share a lOOamino-acid DNA binding domain known as the winged helix motif, which directs monomeric DNA recognition. The winged helix motif is seen by X-ray crystallographic analysis in the form of an a-helical thorax and two wing-like loops, resembling a butterfly (Clark et 1To whom correspondence should be addressed at the Mammalian Genetics Laboratory, ABL-BRP, NCI-FCRDC, P.O. Box B, Bldg. 539, Frederick, MD 21702. Telephone: (301) 846-1260. Fax: (301) 8466666. 0888-7543195 $6.00 Copyright 0 1995 by Academic Press, Inc. All rights of reproduction in any form reserved.
al., 1993). Family members have been isolated from a wide variety of species, including Drosophila, Xenopus, zebrafish, Caenor habditis elegans, yeast, rodents, and humans. These transcription factors play crucial and diverse roles in development and differentiation, as demonstrated by expression patterns and genetic mutations (reviewed by Costa, 1994). These factors have been implicated in the regulation of endoderm differentiation and neuroectoderm formation, as well as morphogenesis of liver and other endoderm-derived organs. Moreover, homologs in different species have been shown to fulfill similar functions (reviewed by Costa, 1994). Clearly, this ancient and well-conserved DNA recognition motif has diverged to fulfill a variety of functional roles. Variations in the DNA-binding domain sequence have been shown to affect the binding specificity of winged helix proteins profoundly (Lai et al., 1991; Overdier et al., 1994). Thus, determining the evolutionary relatedness of different family members is likely to be informative with respect to their regulatory functions in viuo. In this regard, determining the chromosomal location of individual winged helix family members can provide important information about the evolutionary origins and relatedness of different family members (Wilkie et al., 1992). For example, the mapping of Drosophila FKH proteins has revealed tight linkage of several family members, implying that gene duplication events are involved in the expansion of this gene family (Hacker et al., 1992). We have pursued the genetic mapping of eight members of this gene family, HNF-3 fork head homolog 1 (HFH-l), HFH-3, HFH-4, HFH-5, HFH-6, HFH-8, and Brain factor 1 (BF-1) and BF-2, isolated by low-stringency hybridization with winged helix DNA-binding domain probes (Tao and Lai, 1992; Clevidence et al., 1993, 1994; Hatini et al., 1994). The loci have been named HNF-3 fork head homolog 1 (Hfil), Hflt3, Hfh4,
388
MAPPING
OF WINGED
HELIX
Hfi5, H/W Hfh8, Hfi9, and H/hlO, respectively, in accordance with mouse nomenclature to indicate that these genes belong to the same family of homologs of the HNF-3/fork head transcription factors. Previously, additional genes of the hepatocyte nuclear factor 3 family, HNF-3a, HNF-3P, and HNF-37, were mapped to the same interspecific backcross panel (Avraham et al., 1992). These mapping results provide important new information regarding the evolution of this gene family and predict their chromosomal location in humans and their potential association with known mouse mutations.
TRANSCRIPTION
TABLE 1 Loci Mapped in Interspecific
Locus HfhZ Hfh3 Hfh4 Hfh5 Hfh6 Hfh8 Hfh9
HfhlO
MATERIALS
AND METHODS
The HFH-1 probe was a 1.4-kb EcoRI HFH-1 cDNA isolated from rat lung (Clevidence et al., 1993). The HFH-3 probe was a 2.8-kb EcoRI HFH-3 cDNA isolated from human kidney (Clevidence et al., 1993). The HFH-4 probe was a 700-bp EcoRI rat genomic fragment containing the amino terminus of the HFH-4 DNA binding domain (Clevidence et al., 1993). The HFH-5 probe was a 2.0-kb Sac1 rat genomic fragment containing the entire HFH-5 DNA binding domain (Clevidence et al., 1993). The HFHB probe was a 5.5-kb EcoRI rat genomic fragment containing the entire HFH-6 DNA binding domain (Clevidence et al., 1993). The HFH-8 probe was a 650bp NotI-EcoRI mouse lung HFH-8 cDNA containing the carboxyl terminal amino acid residues of HFH-8 (Clevidence et al., 1994). The BF-1 probe was a 1.6-kb SfiI-XhoI mouse cDNA containing the carboxyl terminal amino acid residues of BF-1 (Tao and Lai, 1992). Finally, the BF-2 probe was a 750-bp EcoRI-BamHI mouse cDNA containing the carboxyl terminal amino acid residues of BF-2 (Hatini et al., 1994).
Probes.
Interspecific backcross mapping. Interspecific backcross progeny were generated by mating (C57BL’6J x Mus spretus)F, females and C57BL/6J males as described (Copeland and Jenkins, 1991). A total of 205 Nz mice were used to map the 8 winged helix-containing loci (see text for details). DNA isolation, restriction enzyme digestion, agarose gel electrophoresis, Southern blot transfer, and hybridization were performed essentially as described (Jenkins et al., 1982). All blots were prepared with Hybond-N’ membrane (Amersham). Probes were labeled with [o-32P1dCTP using a random priming labeling kit (Amersham); washing was done to a final stringency of 0.51.0x SSCP, 0.1% SDS, 65°C. The fragments detected by each of the HFH family member probes in restriction enzyme-digested C57BL/6J and M. spretus DNAs are shown in Table 1. The presence or absence of M. spretus-specific fragments was followed in backcross mice. The map locations of several of the marker loci used to position the various Hfh loci on our interspecific backcross have been previously reported. These loci include the fibroblast growth factor receptor 2 (Fgfr2) and 06-methylguanine DNA methyltransferase (Mgmt) on chromosome 7 (Avraham et al., 1994); haptoglobin (Hp), adenine phosphoribosyl transferase (Aprt), and avian musculoaponeurotic fibrosarcoma oncogene homolog (Maf) on chromosome 8 (Ceci et al., 1990; Bae et al., 1994); reticuloendotheliosis oncogene (Rel), adrenergic receptor, a-1 (A&all, protein kinase C, a (Pkca), and proline 4hydroxylase, p polypeptide (P4hb) on chromosome 11 (Buchberg et al., 1989; McKenzie et al., 1993; Morishige et al., 1994); neuroblastoma myc-related oncogene 1 (Nmycl), son of sevenless 2 (Sosd), and spectrin, p-1 (Spnbl) on chromosome 12 (Oettinger et al., 1991; Avraham et al., 1992; Webb et al., 1993); Friend MuLV integration site 1 (FimZ i, bone morphogenetic protein 6 (BmpG), ras p21 protein activator (Rasa), dihydrofolate reductase (Dhfr), and 5-hydroxytryptamine (serotonin) la receptor (Htrla) on chromosome 13 (Dickinson et al., 1990; Justice et al., 1990; Copeland et al., 1992; Wilkie et al., 1993); and CD5 antigen (Cd51 and Fas antigen (Fas) on chromosome 19 (Benovic et al., 1991; Watanabe-Fukunaga et al., 1992). Recombination distances were calculated as described (Green,
389
FACTORS
Locus name HFH-1 HFH-3 HFH-4 HFH-5 HFH-6 HFH-8 BF-1 BF-2
Backcross
Mice M. spretus
Enzyme
C57BL&J fragment size (kb)
fragment size (kb)
SphI BamHI B&I BamHI BamHI PVUII XbaI XbaI
5.6” 9.1 0.7, 0.4 6.2” 9.0” 5.0, 0.9 12.0 5.9
8.0”,’ 3.5 1.8,0.4 s.2n m LZ.9 -11.0 -9.4”
DMajor bands detected. b Restriction fragment sizes underlined fragments typed in the backcross analysis.
indicate
the restriction
1981) using the computer program SPRETUS MADNESS. Gene order was determined by minimizing the number of recombination events required to explain the allele distribution patterns.
RESULTS
AND DISCUSSION
Mapping Results The murine chromosomal locations of eight members of the HFH family of transcription factors were determined by interspecific backcross (IB) analysis. IB progeny were derived from matings of [(C57BL/6J x M. spretus)F1 X C57BW6Jl mice. This interspecific backcross mapping panel has been typed for over 1600 loci that are well distributed among all of the autosomes as well as the X chromosome (Copeland and Jenkins, 1991). C57BL/6J and M. spretus DNAs were digested with several enzymes and analyzed by Southern blot hybridization for informative restriction fragment length polymorphisms (RFLPs) (Table 1) using probes derived from the eight Hfh loci. The mapping results indicate that in most cases the members of the hepatocyte nuclear factor 3/fork head winged helix family of transcription factors have become well dispersed throughout the mouse genome, as the 11 loci (Hfi and Hnf loci) mapped to 7 different mouse autosomes (Fig. 1; Avraham et al., 1992). In 3 of the 4 cases in which 2 loci mapped to the same mouse autosome (Hnf3y and Hfi5 are 43.7 _’ 3.7 CM apart on chromosome 7; Hfi3 and Hfi4 are 47.1 ? 5.4 CM apart on chromosome 11; and H/h1 and H/%10 are 36.8 -+ 3.9 CM apart on chromosome 13), each locus was well separated from the other and mapped to a different region of human homology (Fig. 1; data not shown), suggesting that the loci were not derived by ancient tandem gene duplication events. Only in the case of Hfi9 and Hnf3a are the genes relatively closely linked, lying 5.3 2 2.0 CM apart from one another on chromosome 12 (Fig. 1; data not shown). Predicted Location Chromosomes
of Winged Helix Genes in Human
The large amount of comparative mapping information that has been accumulated between mouse and
390
AVRAHAM
Fiml
ET AL.
6p23-~223
6
Rel
2p13-p12
Adral
Jq32-q34
19
11 f Pkca
17q22q24
FM
lOq25.3q26
f lw37.9.3f
31136, 22f
z f
Cd5
1o/lo7, 9.3 + 28
2.8
llq13
H&6 5/W,
yfk4
1.3
P4h6
3.2 f 1.4
lu123,
9.8 f27 t
17q25
FaS
~
1cqz3-q24.1
t
<
f
HP s/113. 4.4 f 1.9
16q221
2~24.1 3/116, 2.6 + 1.5
27fl63, 16.6 f 29
/L +
5qt3
Jf/kl@ Lw
5qll.Zq13.2
1
ws, (3.1)
w
Rasa
t< 91175, 5.1 f 1.7
161161, 9.9 22.4
6/138, 4.3 k1.7
1 SOS2
14q21 -q22
Spnbl
14q24.1-q24.2
FIG. 1. Murine chromosomal location of eight Hf?z genes. Eight Hfh genes were mapped to mouse chromosomes by interspecific backcross analysis. Partial chromosome linkage maps showing the location of the Hfl genes in relation to linked markers are shown. The number of recombinant Nz animals over the total number of NZ animals typed plus the recombination frequencies expressed as genetic distance in centimorgans (il standard error) is shown for each pair of loci to the left of the chromosome maps. The upper 95% confidence limit of the recombination distance is given in parentheses when no recombinants were found between loci. Gene order was determined by minimizing the number of recombination events required to explain the allele distribution patterns. No double recombinant5 were detected for any of the loci depicted on the maps; therefore, for the sake of brevity, haplotype data have not been included. The positions of loci mapped to human chromosomes are shown to the right of the chromosome maps. References for the map positions of most human loci can be obtained from GDB (Genome Data Base), a computerized database of human linkage information maintained by the William H. Welch Medical Library of The Johns Hopkins University (Baltimore, MD).
MAPPING Helix 1 mF3Alph.a
BF-2 IiFE-6 BF-1 HFE-3 HFE-6 HFE-5 HFB-1 WI-4 ILF RTLF
OF WINGED
Helix 2
HELIX
TRANSCRIPTION
Helix 3
FACTORS Wing 1
391 Wing 2
HAlrPPYsYIsLITMAI*q*-~~LFPWR*~*~FVKVARSPDKP-GKQSY DsalaMnNxY--LRRQKR?Kc! ~.......A......L.S.B.B......~F.~.....~-.......~..L......IP.E.~.-...N....D.E.AD..D..SF--...B....E E........A......L.S.H.B....S;.~F.SIIB.....BIIEES-.......8..L......IP.E.~H.. ..u..S.P.a.Qll..D..SF--...B....R ~...F..~A..M...R.S.E.E...~..EF..~.....E.B.~-.......N..L.Y.....E.~.P.. ..N M D.S.DDV.IG.TTGK...RSTTSR _.._._ . . . ..SA..A...~..D.B....Q...WA.N..F.NXSKAG-.......H..L....8..P.DE.B.-...N _...._D.w..D..&JF-R.KB..m EE.......A..V....SS...E . . . . . . ..FLQBB..FF.~.~-.B..V..8..L.B..I.L~K~R.-...B...IP.A.EE...E.SF--E..ER~.RB . . . ..SA..A....S..B~....Q...W~..F.K~-.......N..L....B..P.~.. ..N M D.a..D..NF--E.W.m _.._._ RE.......A..A...PDSAGGE? . ..A..~YL.GK..FF.~-.B..V..H..L....... L.D.~R.I?I..~..M.N.H.EYT.AD.YF--B..B..~ .Y......AT..C..M.AS~~I . ..A..Ii..T.B.C.F.W. . . . . ..N..L.B..I..E.EK.E.- . ..QF.BIP.QYBEBLLS.AF--KK.W ~S......~Q..~..~..D.Q...~..TH.TKNY....~. . . . . ..N..L.E.I..P..QBE.. . ..F.EIQ.A.sKLmF--EK.PP.m TS.....F&..Y...mS.N.C.PVK...S..L.H . ..FsW-.S..V..N..L.q..Q..E..E.VN.... L.m_.EYKPNLI-M---.KK.m
65% 64% 57% 56% 54% 52% 50% 47% 47% 45%
FIG. 2. Sequence alignment of 12 winged helix DNA-binding domains derived from the HNF-3/fork head (HFH) family. The amino acid sequence (one-letter code) of the HNF-So DNA-binding domain is compared with the BF-2, HFH-6, BF-1, HFH-3, HFH-8, HFH-6, HFH-1, HFH-4, ILF, and HTLF sequences, which are presented in decreasing order of homology (Lai et al., 1991; Clevidence et al., 1993, 1994; Tao and Lai, 1992; Li et al., 1991, 1992a,b). Identical amino acid residues are indicated by dots, conserved residues are in boldface, and nonconservative changes are underlined. On the amino acid sequence, the positions of helices (1 to 3) and wing structures within the winged helix motif are indicated by bars (Clark et al., 1993).
human chromosomes often makes it possible to predict the chromosomal location of a gene in one species based solely upon its mapping in the other species. For example, the mouse Hfil gene maps within a region of human chromosome 6 homology (Fig. 11, strongly suggesting that the human homolog of Hfi1 will map to human chromosome 6. Likewise, Hfi4, Hfi5, Hfi8, and HfilO map to regions of human chromosome 17q, lOq, 16q, and 5q homology, respectively, strongly suggesting that the human homologs of these genes will map to these human chromosome regions as well. One locus, Hfi3, maps between regions of human chromosome 2p and 5q homology (Fig. 1). As yet, it is not possible to predict which human chromosomal region contains the Hfi3 locus. There appears to be some potential linkage between genes, based on the mapping of two human HNF-3/fork head family transcription factors, HTLF and ILF. We predict (see above) that the human homolog of Hfi3 will map to 2p or 5q; HTLF is located on 2p16-~21 (Li et al., 1992b). The different amino acid homology of the DNA binding domains of HFH3 and HTLF to the domain of HNF-3(-U (56 and 45%, respectively) suggests that these two genes are not homologs of one another (Fig. 2). ILF is located on 17q25 (Li et al., 1992a); based on the location of markers flanking Hfi4, we predict that the human homolog will also map to the long arm of chromosome 17 (see above). Although HFH4 and ILF both have 47% amino acid homology to the DNA binding domain of HNF-3a, they differ between themselves in the amino acid sequence of the binding domain and completely diverge outside the winged helix motif (Clevidence et al., 1994), suggesting that they are indeed different genes (Fig. 2). Based on these observations, it is possible that the human homologs for Hfi3 and Hfi4 will be linked to HTLF and ILF, respectively. Whether a potential linkage will provide functional interactions between the genes remains to be determined. Finally, our mapping data strongly support the notion that the rat BF-1 gene is the rodent homolog of the human HFKl gene. The human HFKl cDNA is 91.2% homologous at the nucleotide level to the rat
BFl cDNA, and HFKl was recently mapped to human chromosome 14qll-q13 (Murphy et al., 19941, consistent with our map location of Hfi9 (Fig. 1). The mapping of Hfi9 to mouse chromosome 12 defines the proximal end of the 14q homology unit, which extends distally for the remainder of the chromosome (Fig. 1; data not shown). Linkage of the Winged Helix Genes to Known Mouse Mutations To look for possible linkage of H/h genes with known mouse mutations, we compared our interspecific maps of chromosomes 7, 8, 11, 12, 13, and 19 with the composite mouse linkage map that reports the map location of many uncloned mouse mutations (compiled by M. T. Davisson, T. H. Roderick, A. L. Hillyard, and D. P. Doolittle and provided from GBASE and The Encyclopedia of the Mouse Genome databases maintained at The Jackson Laboratory, Bar Harbor, ME). Descriptions of the phenotypes of mouse mutations mapping near Hfi genes were obtained from Green (1989). Five of the Hfi genes, Hfhl, Hfh5, Hfi6, H/%8, and Hfh9, did not map near a known mouse mutation with a phenotype consistent with what would be predicted for a mutation in one of these genes. However, the expression patterns of many of these genes have been characterized only to a limited extent, and it is still conceivable that a known mouse mutation may result from a defect in one of these Hfh genes. Three Hfi genes map near mouse mutations that could conceivably result from a defect in one of these genes. For example, ames dwarf (df 1, vestigial-tail Cut), wobbler (wr), and chylous ascites (Chy) map to the proximal region of chromosome 11 near the Hfi3 locus. While df, vt, and wr do not appear to be good candidates [df maps distal to Adral (Buckwalter et al., 1991), wr maps proximal to ReZ (Kaupmann et al., 19921, and ut results from a defect in Wnt3a (Camper et al., 199411, the Chy mutation could result from a defect in Hfi3. Chy mice are recognizable by the presence of milky fluid in the abdomen and swollen hind feet resulting from tissue edema. Hfi3 is expressed in the kidney
AVRAHAM
392
(Clevidence et al., 19931, and defects in Hfi3 could thus possibly produce the tissue edema seen in Chy mice. Several mutations lie in the distal region of chromosome 11 near Hfi4, including Jackson shaker (&I, cerebellar outflow degeneration (cod 1, and teetering (tn). js mice exhibit circling and head shaking, which is probably due to an inner ear abnormality; cod mice are lethargic and ataxic, with degenerative changes found in neurons; and tn mice, which die by 6 weeks of age, initially have unstable movements and are growth retarded. Hfi4 is expressed in the brain and lung (Clevidence et al., 1993, 1994), suggesting a potential link between H/%4 and any of these three mouse mutations. Two recessive mutations map in the vicinity of HfilO, furless (fi) and pearl (pe). Furless mice exhibit abnormal hair, with reduced viability prior to weaning. pe mice display several abnormalities, including a reduced ipsilateral projection from the retina to the lateral geniculate nucleus, the superior colliculus, and the visual cortex. HfhlO is expressed during development in the temporal half of the retina (Hatini et al., 19941, suggesting a possible link between HfilO and pe. ACKNOWLEDGMENTS We thank D. J. Gilbert, D. Swing, M. Barnstead, and D. Barnhart for excellent technical assistance. This research was supported, in part, by the National Cancer Institute, DHHS, under Contract NOlCO-74101 with ABL. This research was also supported, in part, by a grant from the Council for Tobacco Research (2822) to R.H.C. and by the American Cancer Society to E.L. K.B.A. is supported by the National Institutes of Health, National Research Service Award 5F32GM15909-02 from the National Institute of General Medical Sciences. R.H.C. is an Established Investiaator of the American Heart Association/Bristol-Myers Squibb.
REFERENCES Avraham, K. B., Givol, D., Avivi, A., Yayon, A., Copeland, N. G., and Jenkins, N. A. (1994). Mapping of murine fibroblast growth factor receptors refines regions of homology between mouse and human chromosomes. Genomics 21: 656-658. Avraham, K. B., Prezioso, V. R., Chen, W. S., Lai, E., Sladek, F. M., Zhong, W., Darnell, J. E., Jr., Jenkins, N. A., and Copeland, N. G. (1992). Murine chromosomal location of four hepatocyte-enriched transcription factors: HNF-So, HNF-30, HNF-37, and HNF-4. Genomics 13: 264-268. Bae, S., Ogawa, E., Maruyama, M., Oda, H., Stake, M., Shigesada, K., Jenkins, N. A., Gilbert, D. J., Copeland, N. G., and Ito, Y. (1994). PEBP2aBlmouse AMLl consists of multiple isoforms that possess differential transactivation potentials. Mol. Cell. Biol. 14: 3242-3252.
ET AL. mosome 526.
11 in an intersubspecific
backcross.
Genomics
10: 515-
Camper, S. A., Greco, T. L., Newhouse, M. M., Takada, S., and McMahon, A. P. (1994). Wnt-3a is critical for caudal embryonic development. Am. J. Hum. Genet. 55: A46. Ceci, J. D., Justice, M. J., Lock, L. F., Jenkins, N. A., and Copeland, N. G. (19901. An interspecific backcross linkage map of mouse chromosome 8. Genomics 6: 72-79. Clark, K., Halay, E., Lai, E., and Burley, S. (1993). Cocrystal structure of the HNF-B/Fork head DNA recognition motif resembles histone H5. Nature 364: 412-420. Clevidence, D. E., Overdier, D. G., Peterson, R. S., Porcella, A., Ye, H., Paulson, E. K., and Costa, R. H. (1994). Members of the HNF3/Forkbead family of transcription factors exhibit distinct cellular expression patterns in lung and regulate the surfactant B promoter. Deu. Biol. 166: 195-209. Clevidence, D. E., Overdier, D. G., Tao, W., Qian, X., Pani, L., Lai, E., and Costa, R. H. (1993). Identification of nine tissue-specific transcription factors of the hepatocyte nuclear factor 3/forkhead DNA-binding-domain family. Proc. Natl. Acad. Sci. USA 90: 39483952. Copeland, N. G., and Jenkins, N. A. (1991). Development and applications of a molecular genetic linkage map of the mouse genome. Trends Genet. 7: 113-118. Copeland, N. G., Gilbert, D. J., Chretien, M., Seidah, N. G., and Jenkins, N. A. (1992). Regional localization of three convertases, PC1 (Net-11, PC2 (Net-21, and furin (Fur), on mouse chromosomes. Genomics 13: 1356-1358. Costa, R. H. (1994). Hepatocyte nuclear factor 3/fork head protein family: Mammalian transcription factors that possess divergent cellular expression patterns and binding specificities. In “Liver Gene Transcription”(F. Tronche and M. Yaniv, Eds.), R. G. Landes, Austin. Texas. Dickinson, M. E., Kobrin, M. S., Silan, C. M., Kingsley, D. M., Justice, M. J., Miller, D. A., Ceci, J. D., Lock, L. F., Lee, A., Buchberg, A. M., Siracusa, L. D., Lyons, K. M., Derynck, R., Hogan, B. L. M., Copeland, N. G., and Jenkins, N. A. (1990). Chromosomal localization of seven members of the murine TGF-fl superfamily ,11.1 . 1 1 1. ,_i,~.” suggests close nnzages to several morpnogenenc mmam 10~1.crenomics 6: 505-520. Green, E. L. (Ed.) (1981). Linkage, recombination, and mapping. In “Genetics and Probability in Animal Breeding Experiments,” pp. 77-113, Macmillan, New York. Green, M. C. (1989). Catalog of mutant genes and polymorphic loci. In “Genetic Variants and Strains of the Laboratory Mouse,” 2nd ed. (M. F. Lyon and A. G. Searle, Eds.), pp. 12-403, Oxford Univ. Press, Oxford. Hacker, U., Grossniklaus, U., Gehring, W. J., and Jackie, H. (1992). Developmentally regulated Drosophila gene family encoding the fork head domain. Proc. Natl. Acad. Sci. USA 89: 8754-8758. Hatini, V., Tao, W., and Lai, E. (1994). Expression of winged helix genes, BF-1 and BF-2, define adjacent domains within the developing forebrain and retina. J. Neurobiol. 25: 1293-1309. Jenkins, N. A., Copeland, N. G., Taylor, B. A., and Lee, B. K. (1982). Organization, distribution and stability of endogenous ecotropic murine leukemia virus DNA sequences in chromosomes of Mus musculus. J. Virol. 43: 26-36.
Benovic, J. L., Onorato, J. J., Arriza, J. L., Stone, W. C., Lohse, M., Jenkins, N. A., Gilbert, D. J., Copeland, N. G., Caron, M. G., and Lefkowitz, R. J. (1991). Cloning, expression and chromosomal localization of o-adrenergic receptor kinase 2: A new member of the receptor kinase family. J. Biol. Chem. 266: 14939-14946.
Justice, M. J., Silan, C. M., Ceci, J. D., Buchberg, A. M., Copeland, N. G., and Jenkins, N. A. (1990). A molecular genetic linkage map of mouse chromosome 13 anchored by the beige (bg) and satin (Sal loci. Genomics 6: 341-351.
Buchberg, A. M., Brownell, E., Nagata, S., Jenkins, N. A., and Copeland, N. G. (1989). A comprehensive genetic map of murine chromosome 11 reveals extensive linkage conservation between mouse and human. Genetics 122: 153-161.
Kaupmann, K., Simon-Chazottes, D., G&net, J.-L., and Jockusch, H. (1992). Wobbler, a mutation affecting motoneuron survival and gonadal functions in the mouse, maps to proximal chromosome 11. Genomics 13: 39-43.
Buckwalter, M. S., Katz, R. W., and Camper, S. A. (1991). Localization of the panhypopituitary dwarf mutation (df) on mouse Chro-
Lai, E., Clark, K. L., Burley, S. K., and Darnell, J. E., Jr. (19931. Hepatocyte nuclear factor 3/fork head or “winged helix” proteins:
MAPPING
OF WINGED
HELIX
A family of transcription factors of diverse biologic function. Proc. N&l. Acad. Sci. USA 90: 10421-10423. Lai, E., Prezioso, V. R., Tao, W., Chen, W. S., and Darnell, J. E., Jr. (1991). Hepatocyte nuclear factor 3o belongs to a gene family in mammals that is homologous to the Drosophila homeotic gene fork head. Genes Dev. 5: 416-427. Li, C., Lai, C., Sigman, D. S., and Gaynor, R. B. (1991). Cloning of a cellular factor, interleukin binding factor, that binds to NFATlike motifs in the human immunodeficiency virus long terminal repeat. Proc. Natl. Acad. Sci. USA 88: 7739-7743. Li, C., Lusis, A. J., Sparkes, R., Nirula, A., and Gaynor, R. (1992a). Characterization and chromosomal mapping of the gene encoding the cellular DNA binding protein ILF. Genomics 13: 665-671. Li, C., Lusis, A. J., Sparkes, R., Tran, S.-M., and Gaynor, R. (1992b). Characterization and chromosomal mapping of the gene encoding the cellular DNA binding protein HTLF. Genomics 13: 658-664. McKenzie, A. N. J., Li, X., Largaespada, D. A., Sato, A., Kaneda, A., Zurawski, S. M., Doyle, E. L., Milatovich, A., Francke, U., Copeland, N. G., Jenkins, N. A., and Zurawshi, G. (1993). Structural comparison and chromosomal localization of the human and mouse IL-13 genes. J. Immunol. 150: 5436-5444.
TRANSCRIPTION
FACTORS
393
Oettinger, H. F., Streeter, H., Lose, E., Copeland, N. G., Gilbert, D. J., Justice, M. J., Jenkins, N. A., Mohandas, T., and Bernfield, M. (1991). Chromosome mapping of the murine syndecan gene. Genomics 11:334-338. Overdier, D. G., Porcella, A., and Costa, R. H. (1994). The DNAbinding specificity of the hepatocyte nuclear factor 3/forkhead domain is influenced by amino acid residues adjacent to the recognition helix. Mol. Cell. Biol. 14: 2755-2766. Tao, W., and Lai, E. (1992). Telencephalon-restricted expression of BF-1, a new member of the HNF-B/fork head gene family, in the developing rat brain, Neuron 8: 957-966. Watanabe-Fukunaga, R., Brannan, C. I., Copeland, N. G., Itoh, N., Yonehara, S., Jenkins, N. A. and Nagata, S. (1992). The cDNA structure, expression and chromosomal assignment of the mouse fas antigen. J. Immunol. 148: 1274-1279. Webb, G. C., Jenkins, N. A., Largaespada, D. A., Copeland, N. G., Fernandez, C. S., and Bowtell, D. D. L. (1993). Mammalian homologues of the Drosophila Son of sevenless gene map to murine chromosomes 17 and 12 and to human chromosomes 2 and 14, respectively. Genomics 18:14-19.
Morishige, K.-I., Takahashi, N., Findlay, I., Koyama, H., Zanelli, J. S., Peterson, C., Jenkins, N. A., Copeland, N. G., Mori, N., and Kurachi, Y. (1993). Molecular cloning, functional expression and localization of an inward rectifier potassium channel in the mouse brain. FEBS Lett. 336: 375-380.
Wilkie, T. M., Gilbert, D. J., Olsen, A. S., Chen, X.-N., Amatruda, T. T., Korenberg, J. R., Trask, B. J., de Jong, P., Reed, R. R., Simon, M. I., Jenkins, N. A., and Copeland, N. G. (1992). Evolution of the mammalian G protein alpha subunit multigene family. Nature Genet. 1:85-91.
Murphy, D. B., Wiese, S., Burfeind, P., Schmundt, D., Mattei, M., Schulz-Schaeffer, W., and Thies, U. (1994). Human brain factor 1, a new member of the fork head gene family. Genomics 21: 551557.
Wilkie, T. M., Chen, Y., Gilbert, D. J., Moore, K. J., Yu, L., Simon, M. I., Copeland, N. G., and Jenkins, N. A. (1993). Identification, chromosomal location, and genome organization of mammalian Gprotein-coupled receptors. Genomics 18:175-184.
25,388-393
(19%)
Murine Chromosomal Location of Eight Members of the Hepatocyte Nuclear Factor 3/Fork Head Winged Helix Family of Transcription Factors KAREN
B. AVRAHAM,* COLIN FLETCHER,* DAVID G. OVERDIER,t DEREK E. CLEVIDENCE,t ROBERT H. CosTA,t NANCY A. JENKINS,* AND NEAL G. COPELAND*,’
ESENG LAI,+
*Mammalian Genetics Laboratory, ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, Frederick, Maryland 2 1702; tDepartment of Biochemistry, College of Medicine, The University of Illinois, Chicago, Illinois 606 12; and *Division of Endocrinology and Cell Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, New York 1002 1 Received August 23, 1994; revised November 5, 1994
A 100.amino-acid DNA-binding motif, known as the winged helix, was first identified in the mammalian hepatocyte nuclear factor-3 (HNF-3) and Drosophila fork head family of transcription factors. Subsequently, more than 40 different genes that contain the winged helix motif have been identified. In the studies described here, we have determined the murine chromosomal location of eight members of this gene family, HFH-1, HFH-3, HFH-4, HFH-5, HFHS, HFH-8, BF-1, and BF-2, by interspecific backcross analysis. These genes, designated HNF-3 fork head homolog 1 (Hfhl), Hfh3, Hfh4, Hfh5, Hfh6, Hfh8, Hfh9, and HfhlO, respectively, mapped to 6 different mouse autosomes and are thus well dispersed throughout the mouse genome. Based on this mapping information, we predict the chromosomal location of these genes in humans and discuss the potential of these genes as candidates for uncloned 0 1996 Academic Press, Inc. mouse mutations.
INTRODUCTION
The study of cell type-specific transcriptional control has led to the identification of a class of transcription factors sharing a DNA-binding domain of the type found in hepatocyte nuclear factor 3 alpha (HNF-3a), HNF-3P, and HNF-3 y and in the Drosophila fork head (FKH) proteins. The nearly 40 genes identified so far (reviewed by Lai et al., 1993; Costa, 1994) share a lOOamino-acid DNA binding domain known as the winged helix motif, which directs monomeric DNA recognition. The winged helix motif is seen by X-ray crystallographic analysis in the form of an a-helical thorax and two wing-like loops, resembling a butterfly (Clark et 1To whom correspondence should be addressed at the Mammalian Genetics Laboratory, ABL-BRP, NCI-FCRDC, P.O. Box B, Bldg. 539, Frederick, MD 21702. Telephone: (301) 846-1260. Fax: (301) 8466666. 0888-7543195 $6.00 Copyright 0 1995 by Academic Press, Inc. All rights of reproduction in any form reserved.
al., 1993). Family members have been isolated from a wide variety of species, including Drosophila, Xenopus, zebrafish, Caenor habditis elegans, yeast, rodents, and humans. These transcription factors play crucial and diverse roles in development and differentiation, as demonstrated by expression patterns and genetic mutations (reviewed by Costa, 1994). These factors have been implicated in the regulation of endoderm differentiation and neuroectoderm formation, as well as morphogenesis of liver and other endoderm-derived organs. Moreover, homologs in different species have been shown to fulfill similar functions (reviewed by Costa, 1994). Clearly, this ancient and well-conserved DNA recognition motif has diverged to fulfill a variety of functional roles. Variations in the DNA-binding domain sequence have been shown to affect the binding specificity of winged helix proteins profoundly (Lai et al., 1991; Overdier et al., 1994). Thus, determining the evolutionary relatedness of different family members is likely to be informative with respect to their regulatory functions in viuo. In this regard, determining the chromosomal location of individual winged helix family members can provide important information about the evolutionary origins and relatedness of different family members (Wilkie et al., 1992). For example, the mapping of Drosophila FKH proteins has revealed tight linkage of several family members, implying that gene duplication events are involved in the expansion of this gene family (Hacker et al., 1992). We have pursued the genetic mapping of eight members of this gene family, HNF-3 fork head homolog 1 (HFH-l), HFH-3, HFH-4, HFH-5, HFH-6, HFH-8, and Brain factor 1 (BF-1) and BF-2, isolated by low-stringency hybridization with winged helix DNA-binding domain probes (Tao and Lai, 1992; Clevidence et al., 1993, 1994; Hatini et al., 1994). The loci have been named HNF-3 fork head homolog 1 (Hfil), Hflt3, Hfh4,
388
MAPPING
OF WINGED
HELIX
Hfi5, H/W Hfh8, Hfi9, and H/hlO, respectively, in accordance with mouse nomenclature to indicate that these genes belong to the same family of homologs of the HNF-3/fork head transcription factors. Previously, additional genes of the hepatocyte nuclear factor 3 family, HNF-3a, HNF-3P, and HNF-37, were mapped to the same interspecific backcross panel (Avraham et al., 1992). These mapping results provide important new information regarding the evolution of this gene family and predict their chromosomal location in humans and their potential association with known mouse mutations.
TRANSCRIPTION
TABLE 1 Loci Mapped in Interspecific
Locus HfhZ Hfh3 Hfh4 Hfh5 Hfh6 Hfh8 Hfh9
HfhlO
MATERIALS
AND METHODS
The HFH-1 probe was a 1.4-kb EcoRI HFH-1 cDNA isolated from rat lung (Clevidence et al., 1993). The HFH-3 probe was a 2.8-kb EcoRI HFH-3 cDNA isolated from human kidney (Clevidence et al., 1993). The HFH-4 probe was a 700-bp EcoRI rat genomic fragment containing the amino terminus of the HFH-4 DNA binding domain (Clevidence et al., 1993). The HFH-5 probe was a 2.0-kb Sac1 rat genomic fragment containing the entire HFH-5 DNA binding domain (Clevidence et al., 1993). The HFHB probe was a 5.5-kb EcoRI rat genomic fragment containing the entire HFH-6 DNA binding domain (Clevidence et al., 1993). The HFH-8 probe was a 650bp NotI-EcoRI mouse lung HFH-8 cDNA containing the carboxyl terminal amino acid residues of HFH-8 (Clevidence et al., 1994). The BF-1 probe was a 1.6-kb SfiI-XhoI mouse cDNA containing the carboxyl terminal amino acid residues of BF-1 (Tao and Lai, 1992). Finally, the BF-2 probe was a 750-bp EcoRI-BamHI mouse cDNA containing the carboxyl terminal amino acid residues of BF-2 (Hatini et al., 1994).
Probes.
Interspecific backcross mapping. Interspecific backcross progeny were generated by mating (C57BL’6J x Mus spretus)F, females and C57BL/6J males as described (Copeland and Jenkins, 1991). A total of 205 Nz mice were used to map the 8 winged helix-containing loci (see text for details). DNA isolation, restriction enzyme digestion, agarose gel electrophoresis, Southern blot transfer, and hybridization were performed essentially as described (Jenkins et al., 1982). All blots were prepared with Hybond-N’ membrane (Amersham). Probes were labeled with [o-32P1dCTP using a random priming labeling kit (Amersham); washing was done to a final stringency of 0.51.0x SSCP, 0.1% SDS, 65°C. The fragments detected by each of the HFH family member probes in restriction enzyme-digested C57BL/6J and M. spretus DNAs are shown in Table 1. The presence or absence of M. spretus-specific fragments was followed in backcross mice. The map locations of several of the marker loci used to position the various Hfh loci on our interspecific backcross have been previously reported. These loci include the fibroblast growth factor receptor 2 (Fgfr2) and 06-methylguanine DNA methyltransferase (Mgmt) on chromosome 7 (Avraham et al., 1994); haptoglobin (Hp), adenine phosphoribosyl transferase (Aprt), and avian musculoaponeurotic fibrosarcoma oncogene homolog (Maf) on chromosome 8 (Ceci et al., 1990; Bae et al., 1994); reticuloendotheliosis oncogene (Rel), adrenergic receptor, a-1 (A&all, protein kinase C, a (Pkca), and proline 4hydroxylase, p polypeptide (P4hb) on chromosome 11 (Buchberg et al., 1989; McKenzie et al., 1993; Morishige et al., 1994); neuroblastoma myc-related oncogene 1 (Nmycl), son of sevenless 2 (Sosd), and spectrin, p-1 (Spnbl) on chromosome 12 (Oettinger et al., 1991; Avraham et al., 1992; Webb et al., 1993); Friend MuLV integration site 1 (FimZ i, bone morphogenetic protein 6 (BmpG), ras p21 protein activator (Rasa), dihydrofolate reductase (Dhfr), and 5-hydroxytryptamine (serotonin) la receptor (Htrla) on chromosome 13 (Dickinson et al., 1990; Justice et al., 1990; Copeland et al., 1992; Wilkie et al., 1993); and CD5 antigen (Cd51 and Fas antigen (Fas) on chromosome 19 (Benovic et al., 1991; Watanabe-Fukunaga et al., 1992). Recombination distances were calculated as described (Green,
389
FACTORS
Locus name HFH-1 HFH-3 HFH-4 HFH-5 HFH-6 HFH-8 BF-1 BF-2
Backcross
Mice M. spretus
Enzyme
C57BL&J fragment size (kb)
fragment size (kb)
SphI BamHI B&I BamHI BamHI PVUII XbaI XbaI
5.6” 9.1 0.7, 0.4 6.2” 9.0” 5.0, 0.9 12.0 5.9
8.0”,’ 3.5 1.8,0.4 s.2n m LZ.9 -11.0 -9.4”
DMajor bands detected. b Restriction fragment sizes underlined fragments typed in the backcross analysis.
indicate
the restriction
1981) using the computer program SPRETUS MADNESS. Gene order was determined by minimizing the number of recombination events required to explain the allele distribution patterns.
RESULTS
AND DISCUSSION
Mapping Results The murine chromosomal locations of eight members of the HFH family of transcription factors were determined by interspecific backcross (IB) analysis. IB progeny were derived from matings of [(C57BL/6J x M. spretus)F1 X C57BW6Jl mice. This interspecific backcross mapping panel has been typed for over 1600 loci that are well distributed among all of the autosomes as well as the X chromosome (Copeland and Jenkins, 1991). C57BL/6J and M. spretus DNAs were digested with several enzymes and analyzed by Southern blot hybridization for informative restriction fragment length polymorphisms (RFLPs) (Table 1) using probes derived from the eight Hfh loci. The mapping results indicate that in most cases the members of the hepatocyte nuclear factor 3/fork head winged helix family of transcription factors have become well dispersed throughout the mouse genome, as the 11 loci (Hfi and Hnf loci) mapped to 7 different mouse autosomes (Fig. 1; Avraham et al., 1992). In 3 of the 4 cases in which 2 loci mapped to the same mouse autosome (Hnf3y and Hfi5 are 43.7 _’ 3.7 CM apart on chromosome 7; Hfi3 and Hfi4 are 47.1 ? 5.4 CM apart on chromosome 11; and H/h1 and H/%10 are 36.8 -+ 3.9 CM apart on chromosome 13), each locus was well separated from the other and mapped to a different region of human homology (Fig. 1; data not shown), suggesting that the loci were not derived by ancient tandem gene duplication events. Only in the case of Hfi9 and Hnf3a are the genes relatively closely linked, lying 5.3 2 2.0 CM apart from one another on chromosome 12 (Fig. 1; data not shown). Predicted Location Chromosomes
of Winged Helix Genes in Human
The large amount of comparative mapping information that has been accumulated between mouse and
390
AVRAHAM
Fiml
ET AL.
6p23-~223
6
Rel
2p13-p12
Adral
Jq32-q34
19
11 f Pkca
17q22q24
FM
lOq25.3q26
f lw37.9.3f
31136, 22f
z f
Cd5
1o/lo7, 9.3 + 28
2.8
llq13
H&6 5/W,
yfk4
1.3
P4h6
3.2 f 1.4
lu123,
9.8 f27 t
17q25
FaS
~
1cqz3-q24.1
t
<
f
HP s/113. 4.4 f 1.9
16q221
2~24.1 3/116, 2.6 + 1.5
27fl63, 16.6 f 29
/L +
5qt3
Jf/kl@ Lw
5qll.Zq13.2
1
ws, (3.1)
w
Rasa
t< 91175, 5.1 f 1.7
161161, 9.9 22.4
6/138, 4.3 k1.7
1 SOS2
14q21 -q22
Spnbl
14q24.1-q24.2
FIG. 1. Murine chromosomal location of eight Hf?z genes. Eight Hfh genes were mapped to mouse chromosomes by interspecific backcross analysis. Partial chromosome linkage maps showing the location of the Hfl genes in relation to linked markers are shown. The number of recombinant Nz animals over the total number of NZ animals typed plus the recombination frequencies expressed as genetic distance in centimorgans (il standard error) is shown for each pair of loci to the left of the chromosome maps. The upper 95% confidence limit of the recombination distance is given in parentheses when no recombinants were found between loci. Gene order was determined by minimizing the number of recombination events required to explain the allele distribution patterns. No double recombinant5 were detected for any of the loci depicted on the maps; therefore, for the sake of brevity, haplotype data have not been included. The positions of loci mapped to human chromosomes are shown to the right of the chromosome maps. References for the map positions of most human loci can be obtained from GDB (Genome Data Base), a computerized database of human linkage information maintained by the William H. Welch Medical Library of The Johns Hopkins University (Baltimore, MD).
MAPPING Helix 1 mF3Alph.a
BF-2 IiFE-6 BF-1 HFE-3 HFE-6 HFE-5 HFB-1 WI-4 ILF RTLF
OF WINGED
Helix 2
HELIX
TRANSCRIPTION
Helix 3
FACTORS Wing 1
391 Wing 2
HAlrPPYsYIsLITMAI*q*-~~LFPWR*~*~FVKVARSPDKP-GKQSY DsalaMnNxY--LRRQKR?Kc! ~.......A......L.S.B.B......~F.~.....~-.......~..L......IP.E.~.-...N....D.E.AD..D..SF--...B....E E........A......L.S.H.B....S;.~F.SIIB.....BIIEES-.......8..L......IP.E.~H.. ..u..S.P.a.Qll..D..SF--...B....R ~...F..~A..M...R.S.E.E...~..EF..~.....E.B.~-.......N..L.Y.....E.~.P.. ..N M D.S.DDV.IG.TTGK...RSTTSR _.._._ . . . ..SA..A...~..D.B....Q...WA.N..F.NXSKAG-.......H..L....8..P.DE.B.-...N _...._D.w..D..&JF-R.KB..m EE.......A..V....SS...E . . . . . . ..FLQBB..FF.~.~-.B..V..8..L.B..I.L~K~R.-...B...IP.A.EE...E.SF--E..ER~.RB . . . ..SA..A....S..B~....Q...W~..F.K~-.......N..L....B..P.~.. ..N M D.a..D..NF--E.W.m _.._._ RE.......A..A...PDSAGGE? . ..A..~YL.GK..FF.~-.B..V..H..L....... L.D.~R.I?I..~..M.N.H.EYT.AD.YF--B..B..~ .Y......AT..C..M.AS~~I . ..A..Ii..T.B.C.F.W. . . . . ..N..L.B..I..E.EK.E.- . ..QF.BIP.QYBEBLLS.AF--KK.W ~S......~Q..~..~..D.Q...~..TH.TKNY....~. . . . . ..N..L.E.I..P..QBE.. . ..F.EIQ.A.sKLmF--EK.PP.m TS.....F&..Y...mS.N.C.PVK...S..L.H . ..FsW-.S..V..N..L.q..Q..E..E.VN.... L.m_.EYKPNLI-M---.KK.m
65% 64% 57% 56% 54% 52% 50% 47% 47% 45%
FIG. 2. Sequence alignment of 12 winged helix DNA-binding domains derived from the HNF-3/fork head (HFH) family. The amino acid sequence (one-letter code) of the HNF-So DNA-binding domain is compared with the BF-2, HFH-6, BF-1, HFH-3, HFH-8, HFH-6, HFH-1, HFH-4, ILF, and HTLF sequences, which are presented in decreasing order of homology (Lai et al., 1991; Clevidence et al., 1993, 1994; Tao and Lai, 1992; Li et al., 1991, 1992a,b). Identical amino acid residues are indicated by dots, conserved residues are in boldface, and nonconservative changes are underlined. On the amino acid sequence, the positions of helices (1 to 3) and wing structures within the winged helix motif are indicated by bars (Clark et al., 1993).
human chromosomes often makes it possible to predict the chromosomal location of a gene in one species based solely upon its mapping in the other species. For example, the mouse Hfil gene maps within a region of human chromosome 6 homology (Fig. 11, strongly suggesting that the human homolog of Hfi1 will map to human chromosome 6. Likewise, Hfi4, Hfi5, Hfi8, and HfilO map to regions of human chromosome 17q, lOq, 16q, and 5q homology, respectively, strongly suggesting that the human homologs of these genes will map to these human chromosome regions as well. One locus, Hfi3, maps between regions of human chromosome 2p and 5q homology (Fig. 1). As yet, it is not possible to predict which human chromosomal region contains the Hfi3 locus. There appears to be some potential linkage between genes, based on the mapping of two human HNF-3/fork head family transcription factors, HTLF and ILF. We predict (see above) that the human homolog of Hfi3 will map to 2p or 5q; HTLF is located on 2p16-~21 (Li et al., 1992b). The different amino acid homology of the DNA binding domains of HFH3 and HTLF to the domain of HNF-3(-U (56 and 45%, respectively) suggests that these two genes are not homologs of one another (Fig. 2). ILF is located on 17q25 (Li et al., 1992a); based on the location of markers flanking Hfi4, we predict that the human homolog will also map to the long arm of chromosome 17 (see above). Although HFH4 and ILF both have 47% amino acid homology to the DNA binding domain of HNF-3a, they differ between themselves in the amino acid sequence of the binding domain and completely diverge outside the winged helix motif (Clevidence et al., 1994), suggesting that they are indeed different genes (Fig. 2). Based on these observations, it is possible that the human homologs for Hfi3 and Hfi4 will be linked to HTLF and ILF, respectively. Whether a potential linkage will provide functional interactions between the genes remains to be determined. Finally, our mapping data strongly support the notion that the rat BF-1 gene is the rodent homolog of the human HFKl gene. The human HFKl cDNA is 91.2% homologous at the nucleotide level to the rat
BFl cDNA, and HFKl was recently mapped to human chromosome 14qll-q13 (Murphy et al., 19941, consistent with our map location of Hfi9 (Fig. 1). The mapping of Hfi9 to mouse chromosome 12 defines the proximal end of the 14q homology unit, which extends distally for the remainder of the chromosome (Fig. 1; data not shown). Linkage of the Winged Helix Genes to Known Mouse Mutations To look for possible linkage of H/h genes with known mouse mutations, we compared our interspecific maps of chromosomes 7, 8, 11, 12, 13, and 19 with the composite mouse linkage map that reports the map location of many uncloned mouse mutations (compiled by M. T. Davisson, T. H. Roderick, A. L. Hillyard, and D. P. Doolittle and provided from GBASE and The Encyclopedia of the Mouse Genome databases maintained at The Jackson Laboratory, Bar Harbor, ME). Descriptions of the phenotypes of mouse mutations mapping near Hfi genes were obtained from Green (1989). Five of the Hfi genes, Hfhl, Hfh5, Hfi6, H/%8, and Hfh9, did not map near a known mouse mutation with a phenotype consistent with what would be predicted for a mutation in one of these genes. However, the expression patterns of many of these genes have been characterized only to a limited extent, and it is still conceivable that a known mouse mutation may result from a defect in one of these Hfh genes. Three Hfi genes map near mouse mutations that could conceivably result from a defect in one of these genes. For example, ames dwarf (df 1, vestigial-tail Cut), wobbler (wr), and chylous ascites (Chy) map to the proximal region of chromosome 11 near the Hfi3 locus. While df, vt, and wr do not appear to be good candidates [df maps distal to Adral (Buckwalter et al., 1991), wr maps proximal to ReZ (Kaupmann et al., 19921, and ut results from a defect in Wnt3a (Camper et al., 199411, the Chy mutation could result from a defect in Hfi3. Chy mice are recognizable by the presence of milky fluid in the abdomen and swollen hind feet resulting from tissue edema. Hfi3 is expressed in the kidney
AVRAHAM
392
(Clevidence et al., 19931, and defects in Hfi3 could thus possibly produce the tissue edema seen in Chy mice. Several mutations lie in the distal region of chromosome 11 near Hfi4, including Jackson shaker (&I, cerebellar outflow degeneration (cod 1, and teetering (tn). js mice exhibit circling and head shaking, which is probably due to an inner ear abnormality; cod mice are lethargic and ataxic, with degenerative changes found in neurons; and tn mice, which die by 6 weeks of age, initially have unstable movements and are growth retarded. Hfi4 is expressed in the brain and lung (Clevidence et al., 1993, 1994), suggesting a potential link between H/%4 and any of these three mouse mutations. Two recessive mutations map in the vicinity of HfilO, furless (fi) and pearl (pe). Furless mice exhibit abnormal hair, with reduced viability prior to weaning. pe mice display several abnormalities, including a reduced ipsilateral projection from the retina to the lateral geniculate nucleus, the superior colliculus, and the visual cortex. HfhlO is expressed during development in the temporal half of the retina (Hatini et al., 19941, suggesting a possible link between HfilO and pe. ACKNOWLEDGMENTS We thank D. J. Gilbert, D. Swing, M. Barnstead, and D. Barnhart for excellent technical assistance. This research was supported, in part, by the National Cancer Institute, DHHS, under Contract NOlCO-74101 with ABL. This research was also supported, in part, by a grant from the Council for Tobacco Research (2822) to R.H.C. and by the American Cancer Society to E.L. K.B.A. is supported by the National Institutes of Health, National Research Service Award 5F32GM15909-02 from the National Institute of General Medical Sciences. R.H.C. is an Established Investiaator of the American Heart Association/Bristol-Myers Squibb.
REFERENCES Avraham, K. B., Givol, D., Avivi, A., Yayon, A., Copeland, N. G., and Jenkins, N. A. (1994). Mapping of murine fibroblast growth factor receptors refines regions of homology between mouse and human chromosomes. Genomics 21: 656-658. Avraham, K. B., Prezioso, V. R., Chen, W. S., Lai, E., Sladek, F. M., Zhong, W., Darnell, J. E., Jr., Jenkins, N. A., and Copeland, N. G. (1992). Murine chromosomal location of four hepatocyte-enriched transcription factors: HNF-So, HNF-30, HNF-37, and HNF-4. Genomics 13: 264-268. Bae, S., Ogawa, E., Maruyama, M., Oda, H., Stake, M., Shigesada, K., Jenkins, N. A., Gilbert, D. J., Copeland, N. G., and Ito, Y. (1994). PEBP2aBlmouse AMLl consists of multiple isoforms that possess differential transactivation potentials. Mol. Cell. Biol. 14: 3242-3252.
ET AL. mosome 526.
11 in an intersubspecific
backcross.
Genomics
10: 515-
Camper, S. A., Greco, T. L., Newhouse, M. M., Takada, S., and McMahon, A. P. (1994). Wnt-3a is critical for caudal embryonic development. Am. J. Hum. Genet. 55: A46. Ceci, J. D., Justice, M. J., Lock, L. F., Jenkins, N. A., and Copeland, N. G. (19901. An interspecific backcross linkage map of mouse chromosome 8. Genomics 6: 72-79. Clark, K., Halay, E., Lai, E., and Burley, S. (1993). Cocrystal structure of the HNF-B/Fork head DNA recognition motif resembles histone H5. Nature 364: 412-420. Clevidence, D. E., Overdier, D. G., Peterson, R. S., Porcella, A., Ye, H., Paulson, E. K., and Costa, R. H. (1994). Members of the HNF3/Forkbead family of transcription factors exhibit distinct cellular expression patterns in lung and regulate the surfactant B promoter. Deu. Biol. 166: 195-209. Clevidence, D. E., Overdier, D. G., Tao, W., Qian, X., Pani, L., Lai, E., and Costa, R. H. (1993). Identification of nine tissue-specific transcription factors of the hepatocyte nuclear factor 3/forkhead DNA-binding-domain family. Proc. Natl. Acad. Sci. USA 90: 39483952. Copeland, N. G., and Jenkins, N. A. (1991). Development and applications of a molecular genetic linkage map of the mouse genome. Trends Genet. 7: 113-118. Copeland, N. G., Gilbert, D. J., Chretien, M., Seidah, N. G., and Jenkins, N. A. (1992). Regional localization of three convertases, PC1 (Net-11, PC2 (Net-21, and furin (Fur), on mouse chromosomes. Genomics 13: 1356-1358. Costa, R. H. (1994). Hepatocyte nuclear factor 3/fork head protein family: Mammalian transcription factors that possess divergent cellular expression patterns and binding specificities. In “Liver Gene Transcription”(F. Tronche and M. Yaniv, Eds.), R. G. Landes, Austin. Texas. Dickinson, M. E., Kobrin, M. S., Silan, C. M., Kingsley, D. M., Justice, M. J., Miller, D. A., Ceci, J. D., Lock, L. F., Lee, A., Buchberg, A. M., Siracusa, L. D., Lyons, K. M., Derynck, R., Hogan, B. L. M., Copeland, N. G., and Jenkins, N. A. (1990). Chromosomal localization of seven members of the murine TGF-fl superfamily ,11.1 . 1 1 1. ,_i,~.” suggests close nnzages to several morpnogenenc mmam 10~1.crenomics 6: 505-520. Green, E. L. (Ed.) (1981). Linkage, recombination, and mapping. In “Genetics and Probability in Animal Breeding Experiments,” pp. 77-113, Macmillan, New York. Green, M. C. (1989). Catalog of mutant genes and polymorphic loci. In “Genetic Variants and Strains of the Laboratory Mouse,” 2nd ed. (M. F. Lyon and A. G. Searle, Eds.), pp. 12-403, Oxford Univ. Press, Oxford. Hacker, U., Grossniklaus, U., Gehring, W. J., and Jackie, H. (1992). Developmentally regulated Drosophila gene family encoding the fork head domain. Proc. Natl. Acad. Sci. USA 89: 8754-8758. Hatini, V., Tao, W., and Lai, E. (1994). Expression of winged helix genes, BF-1 and BF-2, define adjacent domains within the developing forebrain and retina. J. Neurobiol. 25: 1293-1309. Jenkins, N. A., Copeland, N. G., Taylor, B. A., and Lee, B. K. (1982). Organization, distribution and stability of endogenous ecotropic murine leukemia virus DNA sequences in chromosomes of Mus musculus. J. Virol. 43: 26-36.
Benovic, J. L., Onorato, J. J., Arriza, J. L., Stone, W. C., Lohse, M., Jenkins, N. A., Gilbert, D. J., Copeland, N. G., Caron, M. G., and Lefkowitz, R. J. (1991). Cloning, expression and chromosomal localization of o-adrenergic receptor kinase 2: A new member of the receptor kinase family. J. Biol. Chem. 266: 14939-14946.
Justice, M. J., Silan, C. M., Ceci, J. D., Buchberg, A. M., Copeland, N. G., and Jenkins, N. A. (1990). A molecular genetic linkage map of mouse chromosome 13 anchored by the beige (bg) and satin (Sal loci. Genomics 6: 341-351.
Buchberg, A. M., Brownell, E., Nagata, S., Jenkins, N. A., and Copeland, N. G. (1989). A comprehensive genetic map of murine chromosome 11 reveals extensive linkage conservation between mouse and human. Genetics 122: 153-161.
Kaupmann, K., Simon-Chazottes, D., G&net, J.-L., and Jockusch, H. (1992). Wobbler, a mutation affecting motoneuron survival and gonadal functions in the mouse, maps to proximal chromosome 11. Genomics 13: 39-43.
Buckwalter, M. S., Katz, R. W., and Camper, S. A. (1991). Localization of the panhypopituitary dwarf mutation (df) on mouse Chro-
Lai, E., Clark, K. L., Burley, S. K., and Darnell, J. E., Jr. (19931. Hepatocyte nuclear factor 3/fork head or “winged helix” proteins:
MAPPING
OF WINGED
HELIX
A family of transcription factors of diverse biologic function. Proc. N&l. Acad. Sci. USA 90: 10421-10423. Lai, E., Prezioso, V. R., Tao, W., Chen, W. S., and Darnell, J. E., Jr. (1991). Hepatocyte nuclear factor 3o belongs to a gene family in mammals that is homologous to the Drosophila homeotic gene fork head. Genes Dev. 5: 416-427. Li, C., Lai, C., Sigman, D. S., and Gaynor, R. B. (1991). Cloning of a cellular factor, interleukin binding factor, that binds to NFATlike motifs in the human immunodeficiency virus long terminal repeat. Proc. Natl. Acad. Sci. USA 88: 7739-7743. Li, C., Lusis, A. J., Sparkes, R., Nirula, A., and Gaynor, R. (1992a). Characterization and chromosomal mapping of the gene encoding the cellular DNA binding protein ILF. Genomics 13: 665-671. Li, C., Lusis, A. J., Sparkes, R., Tran, S.-M., and Gaynor, R. (1992b). Characterization and chromosomal mapping of the gene encoding the cellular DNA binding protein HTLF. Genomics 13: 658-664. McKenzie, A. N. J., Li, X., Largaespada, D. A., Sato, A., Kaneda, A., Zurawski, S. M., Doyle, E. L., Milatovich, A., Francke, U., Copeland, N. G., Jenkins, N. A., and Zurawshi, G. (1993). Structural comparison and chromosomal localization of the human and mouse IL-13 genes. J. Immunol. 150: 5436-5444.
TRANSCRIPTION
FACTORS
393
Oettinger, H. F., Streeter, H., Lose, E., Copeland, N. G., Gilbert, D. J., Justice, M. J., Jenkins, N. A., Mohandas, T., and Bernfield, M. (1991). Chromosome mapping of the murine syndecan gene. Genomics 11:334-338. Overdier, D. G., Porcella, A., and Costa, R. H. (1994). The DNAbinding specificity of the hepatocyte nuclear factor 3/forkhead domain is influenced by amino acid residues adjacent to the recognition helix. Mol. Cell. Biol. 14: 2755-2766. Tao, W., and Lai, E. (1992). Telencephalon-restricted expression of BF-1, a new member of the HNF-B/fork head gene family, in the developing rat brain, Neuron 8: 957-966. Watanabe-Fukunaga, R., Brannan, C. I., Copeland, N. G., Itoh, N., Yonehara, S., Jenkins, N. A. and Nagata, S. (1992). The cDNA structure, expression and chromosomal assignment of the mouse fas antigen. J. Immunol. 148: 1274-1279. Webb, G. C., Jenkins, N. A., Largaespada, D. A., Copeland, N. G., Fernandez, C. S., and Bowtell, D. D. L. (1993). Mammalian homologues of the Drosophila Son of sevenless gene map to murine chromosomes 17 and 12 and to human chromosomes 2 and 14, respectively. Genomics 18:14-19.
Morishige, K.-I., Takahashi, N., Findlay, I., Koyama, H., Zanelli, J. S., Peterson, C., Jenkins, N. A., Copeland, N. G., Mori, N., and Kurachi, Y. (1993). Molecular cloning, functional expression and localization of an inward rectifier potassium channel in the mouse brain. FEBS Lett. 336: 375-380.
Wilkie, T. M., Gilbert, D. J., Olsen, A. S., Chen, X.-N., Amatruda, T. T., Korenberg, J. R., Trask, B. J., de Jong, P., Reed, R. R., Simon, M. I., Jenkins, N. A., and Copeland, N. G. (1992). Evolution of the mammalian G protein alpha subunit multigene family. Nature Genet. 1:85-91.
Murphy, D. B., Wiese, S., Burfeind, P., Schmundt, D., Mattei, M., Schulz-Schaeffer, W., and Thies, U. (1994). Human brain factor 1, a new member of the fork head gene family. Genomics 21: 551557.
Wilkie, T. M., Chen, Y., Gilbert, D. J., Moore, K. J., Yu, L., Simon, M. I., Copeland, N. G., and Jenkins, N. A. (1993). Identification, chromosomal location, and genome organization of mammalian Gprotein-coupled receptors. Genomics 18:175-184.