- Email: [email protected]
Chromosomal Localization ofCdx2,a Murine Homologue of theDrosophilaGeneCaudal,to Mouse Chromosome 5
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3.
4.
5.
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BRIEF REPORTS human genomic DNA. Proc. Natl. Acad. Sci. USA 91: 5695– 5699. Groden, J., Thliveris, A., Samowitz, W., Carlson, M., Gelbert, L., Albertsen, H., Joslyn, G., Stevens, J., Spirio, L., Robertson, M., Sargeant, L., Krapcho, K., Wolff, E., Burt, R., Hughes, J. P., Warrington, J., McPherson, J., Wasmuth, J., Le Paslier, D., Abderrahim, H., Cohen, D., Leppert, M., and White, R. (1991). Identification and characterization of the familial adenomatous coli gene. Cell 66: 589–600. Hampton, G. M., Ward, J. R., Cottrell, S., Howe, K., Thomas, H. J., Ballhausen, W. G., Jones, T., Sheer, D., Solomon, E., Frischauf, A. M., and Bodmer, W. (1992). Yeast artificial chromosomes for the molecular analysis of the familial polyposis APC gene region. Proc. Natl. Acad. Sci. USA 89: 8249–8253. Joslyn, G., Carlson, M., Thliveris, A., Albertsen, H., Gelbert, L., Samowitz, W., Groden, J., Stevens, J., Spirio, L., Robertson, M., Krapcho, K., Sargeant, L., Wolff, E., Burt, R., Hughes, J. P., Warrington, J., McPherson, J., Wasmuth, J., Le Paslier, D., Abderrahim, H., Cohen, D., Leppert, M., and White, R. (1991). Identification of deletion mutations and three new genes at the familial polyposis locus. Cell 66: 601–613. Kinzler, K. W., Nilbert, M. C., Su, L. K., Vogelstein, B., Bryan, T. M., Levy, D. B., Smith, K. J., Preisinger, A. M., Hedge, P., McKechnie, D., Finnear, R., Markham, A., Groffen, J., Boguski, M. S., Altschul, S. F., Horii, A., Ando, H., Miyoshi, Y., Miki, Y., Nishisho, I., and Nakamura, Y. (1991). Identification of FAP locus genes from chromosome 5q21. Science 253: 661–665. Leppert, M., Dobbs, M., Scambler, P., O’Connell, P., Nakamura, Y., Stauffer, D., Woodward, S., Burt, R. W., Hughes, J. P., Gardner, E. J., Lathrop, M., Wasmuth, J., Lalouel, J. M., and White, R. (1987). The gene for familial polyposis coli maps to the long arm of chromosome 5. Science 238: 1411–1413.
Chromosomal Localization of Cdx2, a Murine Homologue of the Drosophila Gene Caudal, to Mouse Chromosome 5 Kallayanee Chawengsaksophak and Felix Beck1
have recently been characterized: Cdx1 (2), Cdx2 (6), and Cdx4 (3). These three genes exhibit some overlapping as well as unique tissue-specific expression during embryogenesis. Interestingly, expression in the embryonic gut is conserved in both Drosophila caudal (9) and its mouse homologues: Cdx1, Cdx2, and Cdx4. Cdx1 is located on mouse chromosome 18. Recently, Cdx4 has been located on the mouse X chromosome (5) but Cdx2 has not been located. Here we report the localization of Cdx2 in the mouse genome using interspecific backcross mapping. A DNA panel from interspecific backcrossed mice (C57BL/ 6J 1 Mus spretus) F1 1 Mus spretus (BSS) was obtained from the Jackson Laboratory (11). Restriction fragment length polymorphisms (RFLPs) between the two parental strains were identified by Southern analysis using various restriction enzymes and a Cdx2-specific probe (a BamHI to StyI fragment of 400 bp). Polymorphic bands of 2.3 and 3.8 kb were detected between Mus spretus and C57BL/6J, respectively, with the restriction enzyme BamHI. Additional polymorphisms were seen with HindIII (12 and 17 kb) and with BglII (8 and 13 kb). DNA from 94 backcross animals was typed. The segregation pattern of the backcross animals was sent to the Jackson Laboratory to compare to that of known markers. Cdx2 is linked to a number of loci assigned to the distal end of mouse chromosome 5 (Fig. 1). These loci are D5Mit9 (The Jackson Laboratory Backcross public database, accessible on the World Wide Web at http:/www.jax.org/resources/documents/ cmdata), nitric oxide synthase (Nos1) (7), pancreatic and duodenal homeobox gene 1 (Pdx1), previously known as insulin promoter factor 1 (Ipf1) (8), polytropic murine leukemia virus 12 (Pmv12) (11), and cytoskeletal beta actin (Actb) (11). The most probable order of these loci is D5Mit9, Nos1, (Pdx1, Pmv12, Cdx2), and Actb (Fig. 1). The location of human CDX2 has not been directly assigned. However, human CDX3 was localized to chromosome 13q12.3 using fluorescence in situ hybridization with a Cdx3 probe (4). Cdx3 in hamster is orthologous to Cdx2 in mouse (14), suggesting that human CDX3 is equivalent to CDX2. Our mapping results further support this: mouse Cdx2 is closely linked to PDX1 on chromosome 5 (Fig. 1), a region
Howard Florey Institute of Experimental Physiology and Medicine, The University of Melbourne, Parkville, Victoria, Australia 3052 Received January 18, 1996; accepted March 18, 1996
Members of the homeobox gene family share a conserved sequence of 180 nucleotides encoding 60 amino acids known as the homeodomain. The homeodomain-containing proteins act as transcription factors, which often play significant roles in pattern formation and in determining the positional identity of the cells during development (for reviews see 12). Drosophila caudal is a homeobox-containing gene first described by Mlodzik (10). Three caudal homologues in mouse 1
To whom correspondence should be addressed. Telephone: 001161-03-9344-7282. Fax: 0011-61-03-9348-1707. GENOMICS
FIG. 1. Haplotype figure for the genetic mapping of Cdx2 on the Jackson Laboratory BSS backcross. The loci are listed proximal to distal to the left of the figure. The black boxes represent the C57BL/ 6J allele, and the white boxes represent the SPRET/Ei allele. The numbers of the animals with each haplotype is given at the bottom of each column of boxes. The frequency of recombination (R) and the standard error (SE) for each interval are given to the right of the figure. The Actb locus is the most distal chromosome 5 marker typed to date in this cross.
34, 270–271 (1996) 0286
ARTICLE NO.
0888-7543/96 $18.00 Copyright q 1996 by Academic Press, Inc. All rights of reproduction in any form reserved.
AID
Genom BREP
/
6r16$$$381
05-09-96 12:40:41
gnmbr
AP: Genomics
271
BRIEF REPORTS
that may be syntenic to human chromosome 13q12 since human PDX1 is localized to 13q12.1 (13). The expression of Drosophila caudal and of mouse Cdx genes in the embryonic gut suggests that they may have a conserved role in the development of these structures. In mammals the further expression of Cdx2 in the extraembryonic membrane (1) is understandable if the phylogenetic connection of the chorioallantoic placenta with hindgut is considered. No information exists concerning placental expression of either Cdx1 or Cdx4. Gene ablation studies should help to clarify the functional role of the Cdx gene family in intestinal/ placental development.
5.
6.
7.
8.
ACKNOWLEDGMENTS
9.
The work was supported by a NH&MRC block grant to the Howard Florey Institute. We thank Lucy Rowe (The Jackson Laboratory, USA) for data analysis and valuable discussions and Kerry Fowler (Murdoch Institute, Melbourne, Australia) for critical comments. We thank Dr. Heather Mack for the use of experimental material.
11.
REFERENCES 1. Beck, F., Erler, T., and James, R. (1995). Expression of Cdx-2 in the mouse embryos and placenta: Possible role in patterning of the extraembryonic membranes. Dev. Dyn. 204: 217–229. 2. Duprey, P., Chowdhury, K., Dressler, G. R., Balling, R., Simon, D., Guenet, J.-L., and Gruss, P. (1988). A mouse gene homologous to the Drosophila gene caudal is expressed in epithelial cells from the embryonic intestine. Genes Dev. 2: 1647–1654. 3. Gamer, L., and Wright, C. V. E. (1993). Murine Cdx-4 bears striking similarities to the Drosophila caudal gene in its homeodomain sequence and early expression pattern. Mech. Dev. 43: 71–81. 4. German, M. S., Wang, J., Fernald, A. A., Espinosa, R. III, Le Beau, M. M., and Bell, G. I. (1994). Localisation of the genes
AID
Genom BREP
/
6r16$$181x
10.
12.
13.
14.
05-09-96 12:40:41
encoding two transcription factors, LMX1 and CDX3, regulating insulin gene expression to human chromosomes 1 and 13. Genomics 24: 403–404. Horn, J. M., and Ashworth, A. (1995). A member of the caudal family of homeobox genes maps to the X-inactivation centre region of the mouse and human X chromosome. Hum. Mol. Genet. 4: 1041–1047. James, R., and Kazenwadel, J. (1991). Homeobox gene expression in the intestine epithelium of adult mice. J. Biol. Chem. 266: 3246–3251. Lee, C. G., Gregg, A. R., and O’Brien, W. E. (1995). Localisation of the neuronal nitric oxide synthase to mouse chromosome 5. Mamm. Genome 6: 56–57. Leiter, E. H. (1995). Linkage of insulin promoter factor 1 (ipf1) to Pmv12 and Actb on chromosome 5. Mouse Genome 93: 150. Macdonald, P. M., and Struhl, G. (1986). A molecular gradient in early Drosophila embryos and its role in specifying the body pattern. Nature 324: 537–545. Mlodzik, M., Fjose, A., and Gehring, W. J. (1985). Isolation of caudal, a Drosophila homeobox-containing gene with maternal expression, whose transcripts form a concentration gradient at the pre-blastoderm stage. EMBO J. 4: 2961–2969. Rowe, L. B., Nadeau, J. H., Turner, R., Frankel, W. N., Letts, V. A., Epping, J. T., Ko, M. S. H., Thurston, S. J., and Birkenmeier, E. H. (1994). Maps from two interspecific backcross DNA panels available as a community genetic mapping resource. Mamm. Genome 5: 253–274. Shashikant, C. S., Utset, M. F., Violette, S. M., Wise, T. L., Einat, P., Pendleton, J. W., Schughart, K., and Ruddle, F. H. (1991). Homeobox genes in mouse development. Eukaryotic Gene Expression 1: 207–245. Stoffel, M., Stein, R., Wright, C. V. E., Espinosa, R. III, Le Beau, M. M., and Bell, G. I. (1995). Location of human homeodomain transcription factor insulin promoter factor 1 (IPF1) to chromosome band 13q12.1. Genomics 28: 125–126. Suh, E., Chen, L., Taylor, J., and Traber, P. (1994). A homeodomain protein related to caudal regulates intestine-specific gene transcription. Mol. Cell. Biol. 14: 7340–7351.
gnmbr
AP: Genomics
2.
3.
4.
5.
6.
BRIEF REPORTS human genomic DNA. Proc. Natl. Acad. Sci. USA 91: 5695– 5699. Groden, J., Thliveris, A., Samowitz, W., Carlson, M., Gelbert, L., Albertsen, H., Joslyn, G., Stevens, J., Spirio, L., Robertson, M., Sargeant, L., Krapcho, K., Wolff, E., Burt, R., Hughes, J. P., Warrington, J., McPherson, J., Wasmuth, J., Le Paslier, D., Abderrahim, H., Cohen, D., Leppert, M., and White, R. (1991). Identification and characterization of the familial adenomatous coli gene. Cell 66: 589–600. Hampton, G. M., Ward, J. R., Cottrell, S., Howe, K., Thomas, H. J., Ballhausen, W. G., Jones, T., Sheer, D., Solomon, E., Frischauf, A. M., and Bodmer, W. (1992). Yeast artificial chromosomes for the molecular analysis of the familial polyposis APC gene region. Proc. Natl. Acad. Sci. USA 89: 8249–8253. Joslyn, G., Carlson, M., Thliveris, A., Albertsen, H., Gelbert, L., Samowitz, W., Groden, J., Stevens, J., Spirio, L., Robertson, M., Krapcho, K., Sargeant, L., Wolff, E., Burt, R., Hughes, J. P., Warrington, J., McPherson, J., Wasmuth, J., Le Paslier, D., Abderrahim, H., Cohen, D., Leppert, M., and White, R. (1991). Identification of deletion mutations and three new genes at the familial polyposis locus. Cell 66: 601–613. Kinzler, K. W., Nilbert, M. C., Su, L. K., Vogelstein, B., Bryan, T. M., Levy, D. B., Smith, K. J., Preisinger, A. M., Hedge, P., McKechnie, D., Finnear, R., Markham, A., Groffen, J., Boguski, M. S., Altschul, S. F., Horii, A., Ando, H., Miyoshi, Y., Miki, Y., Nishisho, I., and Nakamura, Y. (1991). Identification of FAP locus genes from chromosome 5q21. Science 253: 661–665. Leppert, M., Dobbs, M., Scambler, P., O’Connell, P., Nakamura, Y., Stauffer, D., Woodward, S., Burt, R. W., Hughes, J. P., Gardner, E. J., Lathrop, M., Wasmuth, J., Lalouel, J. M., and White, R. (1987). The gene for familial polyposis coli maps to the long arm of chromosome 5. Science 238: 1411–1413.
Chromosomal Localization of Cdx2, a Murine Homologue of the Drosophila Gene Caudal, to Mouse Chromosome 5 Kallayanee Chawengsaksophak and Felix Beck1
have recently been characterized: Cdx1 (2), Cdx2 (6), and Cdx4 (3). These three genes exhibit some overlapping as well as unique tissue-specific expression during embryogenesis. Interestingly, expression in the embryonic gut is conserved in both Drosophila caudal (9) and its mouse homologues: Cdx1, Cdx2, and Cdx4. Cdx1 is located on mouse chromosome 18. Recently, Cdx4 has been located on the mouse X chromosome (5) but Cdx2 has not been located. Here we report the localization of Cdx2 in the mouse genome using interspecific backcross mapping. A DNA panel from interspecific backcrossed mice (C57BL/ 6J 1 Mus spretus) F1 1 Mus spretus (BSS) was obtained from the Jackson Laboratory (11). Restriction fragment length polymorphisms (RFLPs) between the two parental strains were identified by Southern analysis using various restriction enzymes and a Cdx2-specific probe (a BamHI to StyI fragment of 400 bp). Polymorphic bands of 2.3 and 3.8 kb were detected between Mus spretus and C57BL/6J, respectively, with the restriction enzyme BamHI. Additional polymorphisms were seen with HindIII (12 and 17 kb) and with BglII (8 and 13 kb). DNA from 94 backcross animals was typed. The segregation pattern of the backcross animals was sent to the Jackson Laboratory to compare to that of known markers. Cdx2 is linked to a number of loci assigned to the distal end of mouse chromosome 5 (Fig. 1). These loci are D5Mit9 (The Jackson Laboratory Backcross public database, accessible on the World Wide Web at http:/www.jax.org/resources/documents/ cmdata), nitric oxide synthase (Nos1) (7), pancreatic and duodenal homeobox gene 1 (Pdx1), previously known as insulin promoter factor 1 (Ipf1) (8), polytropic murine leukemia virus 12 (Pmv12) (11), and cytoskeletal beta actin (Actb) (11). The most probable order of these loci is D5Mit9, Nos1, (Pdx1, Pmv12, Cdx2), and Actb (Fig. 1). The location of human CDX2 has not been directly assigned. However, human CDX3 was localized to chromosome 13q12.3 using fluorescence in situ hybridization with a Cdx3 probe (4). Cdx3 in hamster is orthologous to Cdx2 in mouse (14), suggesting that human CDX3 is equivalent to CDX2. Our mapping results further support this: mouse Cdx2 is closely linked to PDX1 on chromosome 5 (Fig. 1), a region
Howard Florey Institute of Experimental Physiology and Medicine, The University of Melbourne, Parkville, Victoria, Australia 3052 Received January 18, 1996; accepted March 18, 1996
Members of the homeobox gene family share a conserved sequence of 180 nucleotides encoding 60 amino acids known as the homeodomain. The homeodomain-containing proteins act as transcription factors, which often play significant roles in pattern formation and in determining the positional identity of the cells during development (for reviews see 12). Drosophila caudal is a homeobox-containing gene first described by Mlodzik (10). Three caudal homologues in mouse 1
To whom correspondence should be addressed. Telephone: 001161-03-9344-7282. Fax: 0011-61-03-9348-1707. GENOMICS
FIG. 1. Haplotype figure for the genetic mapping of Cdx2 on the Jackson Laboratory BSS backcross. The loci are listed proximal to distal to the left of the figure. The black boxes represent the C57BL/ 6J allele, and the white boxes represent the SPRET/Ei allele. The numbers of the animals with each haplotype is given at the bottom of each column of boxes. The frequency of recombination (R) and the standard error (SE) for each interval are given to the right of the figure. The Actb locus is the most distal chromosome 5 marker typed to date in this cross.
34, 270–271 (1996) 0286
ARTICLE NO.
0888-7543/96 $18.00 Copyright q 1996 by Academic Press, Inc. All rights of reproduction in any form reserved.
AID
Genom BREP
/
6r16$$$381
05-09-96 12:40:41
gnmbr
AP: Genomics
271
BRIEF REPORTS
that may be syntenic to human chromosome 13q12 since human PDX1 is localized to 13q12.1 (13). The expression of Drosophila caudal and of mouse Cdx genes in the embryonic gut suggests that they may have a conserved role in the development of these structures. In mammals the further expression of Cdx2 in the extraembryonic membrane (1) is understandable if the phylogenetic connection of the chorioallantoic placenta with hindgut is considered. No information exists concerning placental expression of either Cdx1 or Cdx4. Gene ablation studies should help to clarify the functional role of the Cdx gene family in intestinal/ placental development.
5.
6.
7.
8.
ACKNOWLEDGMENTS
9.
The work was supported by a NH&MRC block grant to the Howard Florey Institute. We thank Lucy Rowe (The Jackson Laboratory, USA) for data analysis and valuable discussions and Kerry Fowler (Murdoch Institute, Melbourne, Australia) for critical comments. We thank Dr. Heather Mack for the use of experimental material.
11.
REFERENCES 1. Beck, F., Erler, T., and James, R. (1995). Expression of Cdx-2 in the mouse embryos and placenta: Possible role in patterning of the extraembryonic membranes. Dev. Dyn. 204: 217–229. 2. Duprey, P., Chowdhury, K., Dressler, G. R., Balling, R., Simon, D., Guenet, J.-L., and Gruss, P. (1988). A mouse gene homologous to the Drosophila gene caudal is expressed in epithelial cells from the embryonic intestine. Genes Dev. 2: 1647–1654. 3. Gamer, L., and Wright, C. V. E. (1993). Murine Cdx-4 bears striking similarities to the Drosophila caudal gene in its homeodomain sequence and early expression pattern. Mech. Dev. 43: 71–81. 4. German, M. S., Wang, J., Fernald, A. A., Espinosa, R. III, Le Beau, M. M., and Bell, G. I. (1994). Localisation of the genes
AID
Genom BREP
/
6r16$$181x
10.
12.
13.
14.
05-09-96 12:40:41
encoding two transcription factors, LMX1 and CDX3, regulating insulin gene expression to human chromosomes 1 and 13. Genomics 24: 403–404. Horn, J. M., and Ashworth, A. (1995). A member of the caudal family of homeobox genes maps to the X-inactivation centre region of the mouse and human X chromosome. Hum. Mol. Genet. 4: 1041–1047. James, R., and Kazenwadel, J. (1991). Homeobox gene expression in the intestine epithelium of adult mice. J. Biol. Chem. 266: 3246–3251. Lee, C. G., Gregg, A. R., and O’Brien, W. E. (1995). Localisation of the neuronal nitric oxide synthase to mouse chromosome 5. Mamm. Genome 6: 56–57. Leiter, E. H. (1995). Linkage of insulin promoter factor 1 (ipf1) to Pmv12 and Actb on chromosome 5. Mouse Genome 93: 150. Macdonald, P. M., and Struhl, G. (1986). A molecular gradient in early Drosophila embryos and its role in specifying the body pattern. Nature 324: 537–545. Mlodzik, M., Fjose, A., and Gehring, W. J. (1985). Isolation of caudal, a Drosophila homeobox-containing gene with maternal expression, whose transcripts form a concentration gradient at the pre-blastoderm stage. EMBO J. 4: 2961–2969. Rowe, L. B., Nadeau, J. H., Turner, R., Frankel, W. N., Letts, V. A., Epping, J. T., Ko, M. S. H., Thurston, S. J., and Birkenmeier, E. H. (1994). Maps from two interspecific backcross DNA panels available as a community genetic mapping resource. Mamm. Genome 5: 253–274. Shashikant, C. S., Utset, M. F., Violette, S. M., Wise, T. L., Einat, P., Pendleton, J. W., Schughart, K., and Ruddle, F. H. (1991). Homeobox genes in mouse development. Eukaryotic Gene Expression 1: 207–245. Stoffel, M., Stein, R., Wright, C. V. E., Espinosa, R. III, Le Beau, M. M., and Bell, G. I. (1995). Location of human homeodomain transcription factor insulin promoter factor 1 (IPF1) to chromosome band 13q12.1. Genomics 28: 125–126. Suh, E., Chen, L., Taylor, J., and Traber, P. (1994). A homeodomain protein related to caudal regulates intestine-specific gene transcription. Mol. Cell. Biol. 14: 7340–7351.
gnmbr
AP: Genomics