- Email: [email protected]
The gene encoding human plasma carboxypeptidase B (CPB2) resides on chromosome 13
BRIEF REPORTS and growth factors (12) and is altered or deleted in a n u m b e r of h u m a n tumors. Chromosome a r m 5q is the site of the familial adenomatous polyposis (FAP) gene, which causes predisposition to colon cancer (1). This region is m u t a t e d or deleted in colon cancer (11), myelodysplastic syndromes (7), and acute myeloid leukemia (AML) (7). If the Gas-1 gene is located on chromosome arm 5q, loss of the gene by m u t a t i o n or deletion may be i m p o r t a n t in the development of colon cancer and AML. Our current studies are aimed at mapping the h u m a n GAS1 gene, using a h u m a n e D N A probe. ACKNOWLEDGMENTS The Gas-1 eDNA probe was kindly provided by Dr. C. Schneider, Centro Internazionale per L'Ingegneria Genetica e Biotechnologia, Trieste. Tina McCarthy is thanked for help with the figure. These studies were supported by a grant from The National Health and Medical Research Council of Australia. REFERENCES 1.
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6. 7.
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Bodmer, W. F., Bailey, C. J., Bodmer, J., Bussey, H. J. R., Ellis, A., Gorman, P., Lucibello, F. C., Murday, V. A., Rider, H. S., Scrambler, P., Sheer, D., Solomon, E., and Spurt, N. K. (1987). Localization of the gene for familial adenomatous polyposis on chromosome 5. Nature 328: 614-616. Ciccarelli, C., Philipson, L., and Sorrentino, V. (1990). Regulation of expression of growth arrest-specific genes in mouse fibroblasts. Mol. Cell Biol. 10: 1525-1529. Colombo, M. P., Baracetti, P., Schneider, C., and Cairo, G. (1989). Aval RFLP at the gas-1 locus on mouse chromosome 12. Nucleic Acids Res. 17: 5415. Colombo, M. P., Martinotti, A., Howard, T. A., Schneider, C., D'Eustachio, P. D., and Seldin, M. F. (1992). Localization of growth arrest-specific genes on mouse Chromosomes 1, 7, 8, 11, 13, and 16. Mammalian Genome 2: 130-134. Evans, E. P. (1989). Standard normal chromosomes. In "Genetic Variants and Strains of the Laboratory Mouse" (M. F. Lyon and A. G. Searle, Ed.), pp. 576-581, Oxford Univ. Press, Oxford. Justice, M. J., and Stephenson, D. A. (1991). Mouse chromosome 13. Mammalian Genome 1(Special Issue): $205-$220. LeBeau, M. M., Westbrook, C. A., Diaz, M. O., Larson, R. A., Rowley, J. D., Gasson, J. C., Golde, D. W., and Sherr, C. J. (1986). Evidence for the involvement of GM-CSF and FMS in the deletion (5q) in myeloid disorders. Science 231: 984-987. LeBeau, M. M., Epstein, N. D., O'Brien, S. J., Nienhuis, A. W., Yang, Y-C., Clark, S. C., and Rowley, J. D. (1987). The interleukin 3 gene is located on human chromosome 5 and is deleted in myeloid leukemias with a deletion of 5q. Proc. Natl. Acad. Sci. USA 84: 5913-5917. Nesbitt, M. N., and Francke, U. (1973). A system of nomenclature for band patterns of mouse chromosomes. Chromosoma 4 1 : 145-158. Schneider, C., Kingl R. M., and Phillipson, L. (1988). Genes specifically expressed at growth arrest of mammalian cells. Cell 54: 787-793. Solomon, E., Voss, R., Hall, V., Bodmer, W. F., Jass, J. R., Jeffreys, A. J., Lucibello, F. C., Patel, I., and Rider, S. H. (1987). Chromosome 5 allele loss in human colorectal carcinomas. Nature 328: 616-619. Wasmuth, J. J., Park, C., and Ferrell, R. E. (1989). Human Gene Mapping 10: Tenth International Workshop on Human Gene Mapping. Cytogenet. Cell. Genet. 51: 137-148.
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14.
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Webb, G. C., Lee, J. S., Campbell, H. D., and Young, I. G. (1989). The genes for interleukins 3 and 5 map to the same locus on mouse chromosome 11. Cytogenet. Cell. Genet. 50: 107-110. Webb, G. C., Campbell, H. D., Lee, J. S., and Young, I. G. (1990). Mapping the gene for murine T-cell growth factor, IL-2, to bands B-C on chromosome 3 and for the a chain of the IL-2 receptor, IL-2ra, to bands A2-A3 on Chromosome 2. Cytogenet. Cell. Genet. 54: 164-168.
The Gene Encoding Human Plasma Carboxypeptidase B (CPB2) Resides on Chromosome 13 SIAO PING TSAI AND DENNIS DRAYNA1 Department of Molecular Biology, Genentech, Inc., 460 Pt. San Bruno Boulevard, South San Francisco, California 94080 Received February 25, 1992; revised July 13, 1992
H u m a n plasma carboxypeptidase B (CPB2) is a recently described carboxypeptidase, first isolated on the basis of its ability to bind plasminogen (4). Although the normal function of this protein is not well understood, it has several characteristics t h a t suggest t h a t it plays a role quite different from t h a t of the known pancreatic carboxypeptidase B (CPB1) (1). We undertook a study to map the C P B 2 gene and to compare its chromosomal location to t h a t of other carboxypeptidases. We used N I G M S hybrid cell mapping panel 2 in our study (3). This mapping panel contains 24 hybrid cell lines. In most cases these lines contain a single h u m a n chromosome on the rodent background, which is either mouse or hamster. We used P C R to determine the presence of the plasma carboxypeptidase B gene in the ~cell lines. We used P C R primers with the following sequences: primer P B P . P C R . 1 3 5' A T G AAG C T T T G C AGC C T T GCA 3' primer P B P . P C R . 1 4 5' CGC GAA GAC A T G C T G C T C ACA 3'. Th ese primers amplify a sequence within the region encoding the hydrophobic leader peptide of CPB2, and the expected P C R product is 63 bp in length. T h e CPB2-specific sequence was amplified using 30 cycles of 94°C for 1 min, 56°C for 2 min, and 72°C for 1.5 min, in reactions containing 0.5 #g of genomic DNA. Th e resultant products were separated on a 6% polyacrylamide gel and visualized by staining with ethidium bromide. T h e results of the P C R reactions using the N I G M S panel 2 D N A s are shown in Fig. 1. Our P C R primers did not amplify 1 To whom correspondence should be addressed. GENOMICS 14, 549-550 (1992) 0888-7543/92 $5.00 Copyright © 1992 by AcademicPress, Inc. All rights of reproduction in any form reserved.
550
BRIEF REPORTS
FIG. 1. Human plasma carboxypeptidase B-specific PCR products. PCR reactions and analysis of the PCR products were performed as described above. Lanes are labeled as follows: HU, HA, and MS are the parental human, hamster, and mouse DNAs, respectively. The numbers indicate the human chromosomes present in each hybrid DNA.
any specific sequence in either mouse or hamster DNA. Only a single hybrid cell line, GM/NA10898, produced an authentic 63-bp CPB2 product. This line contains chromosome 13 as its sole h u m a n component., This result, which is in agreement with results obtained from other somatic cell hybrid mapping panels including N I G M S hybrid mapping panel 1 and a commercially available hybrid panel (2, 3; results not shown), clearly indicates t h a t the C P B 2 coding sequence lies on chromosome 13. Two other cell lines, G M / N A 1 1 4 1 8 and G M / NA06318B, produced a smaller P C R product, approximately 45 bp in length. W h e t h e r these products represent an artifact or amplification of a related sequence on other chromosomes is not known. T h e assignment of C P B 2 to chromosome 13 contrasts this gene with those of other carboxypeptidases. For example, the h u m a n pancreatic carboxypeptidase A gene (CPA1) is known to reside on chromosome 7 (6), while the dipeptidyl carboxypeptidase 1 (angiotensin converting enzyme) gene resides on chromosome 17 (5). These results suggest t h a t the members of the carboxypeptidase gene family reside on many different chromosomes, analogous to those of the serine protease family (1).
REFERENCES 1.
Barrett, A. J., and MacDonald, J. K. (1986). "Mammalian Proteases, a Glossary and Bibliography," Vol. 2, pp. 155-224, Academic Press, London. 2. BIOS Corp., 291 Whitney Ave., New Haven, CT 06511. 3. Coriell Institute for Medical Research, 401 Haddon Ave., Camden, NJ 08103. 4. Eaton, D. L., Malloy, B. E., Tsai, S.-P., Henzel, W., and Drayna, D: (1991). Isolation, molecular cloning, and partial characterization of a novel carboxypeptidase B from human plasma. J. Biol. Chem. 266: 21833-21838. 5. Mattei, M. G., Hubert~ C., Alhenc-Gelas, F., Roeckel, N., Corvol, P., and Soubrier, F. (1989). The angioteusin-I converting enzyme gene is on chromosome 17. Cytogenet. Cell Genet. 51: 1041. 6. Stewart, E. A., Craik, C. S., Hake, L., and Bowcock, A. M. (1990). Human carboxypeptidase A identifies a BglII RFLP and maps to 7q31-qter. Am. J. Hum. Genes. 46: 795-800.
Assignment of the Human Gene Propionyl Coenzyme A Carboxylase, a-Chain, (PCCA) to Chromosome 13q32 by in Situ Hybridization Ingo Kennerknecht, 1 Christine Klett, and Horst Hameister Abtei/ung Klinische Genetik der Universit~t, Parkstrasse 1 I, W-7900 U/m, Germany
Received February 26, 1992
Propionic acidemia is an autosomal recessively inherited disorder of organic acid metabolism caused by deficiency of propionyl-CoA carboxylase [PCC, propanoyl-CoA: carbondioxide ligase (ADP-forming), EC 6.4.1.3]. The native enzyme is a large complex t h a t consists of two nonidentical subunits, most probably six a-subunits and six fl-subunits (6). c D N A clones coding for the a- and B-polypeptides of human PCC were isolated. By studying the segregation of D N A sequences homologous to the PCC cDNAs in a panel of m o u s e - h u m a n hybrids, the gene coding for the a-chain was mapped to chromosome 13 and t h a t for the B-chain was mapped to chromosome 3 (5). F u r t h e r refinement of localization of P C C A to 13q22--~q34 was possible by evaluation of a contiguous genetic linkage map of chromosome 13 (1). The P C C B gene was further regionalized to chromosome 3q13.3--~q22 by in situ hybridization (3). More recently, we have been able to tentatively assign the locus for the a-chain gene to 13q32 by gene dosage studies in cell lines with different chromosomal aberrations of chromosome 13 (2). We have now confirmed this assignment directly by use of a c D N A probe and in situ hybridization to metaphase chromosomes. For this purpose, chromosomes were prepared from P H A stimulated lymphocytes labeled with 20 ~g/pl bromo-desoxyuridine according to standard techniques. The recombinant a To whom correspondence should be addressed. GENOMICS 14, 550--551 (1992) 0888-7543/92 $5.00 Copyright © 1992 by AcademicPress, Inc. All rights of reproduction in any form reserved.
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6. 7.
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Bodmer, W. F., Bailey, C. J., Bodmer, J., Bussey, H. J. R., Ellis, A., Gorman, P., Lucibello, F. C., Murday, V. A., Rider, H. S., Scrambler, P., Sheer, D., Solomon, E., and Spurt, N. K. (1987). Localization of the gene for familial adenomatous polyposis on chromosome 5. Nature 328: 614-616. Ciccarelli, C., Philipson, L., and Sorrentino, V. (1990). Regulation of expression of growth arrest-specific genes in mouse fibroblasts. Mol. Cell Biol. 10: 1525-1529. Colombo, M. P., Baracetti, P., Schneider, C., and Cairo, G. (1989). Aval RFLP at the gas-1 locus on mouse chromosome 12. Nucleic Acids Res. 17: 5415. Colombo, M. P., Martinotti, A., Howard, T. A., Schneider, C., D'Eustachio, P. D., and Seldin, M. F. (1992). Localization of growth arrest-specific genes on mouse Chromosomes 1, 7, 8, 11, 13, and 16. Mammalian Genome 2: 130-134. Evans, E. P. (1989). Standard normal chromosomes. In "Genetic Variants and Strains of the Laboratory Mouse" (M. F. Lyon and A. G. Searle, Ed.), pp. 576-581, Oxford Univ. Press, Oxford. Justice, M. J., and Stephenson, D. A. (1991). Mouse chromosome 13. Mammalian Genome 1(Special Issue): $205-$220. LeBeau, M. M., Westbrook, C. A., Diaz, M. O., Larson, R. A., Rowley, J. D., Gasson, J. C., Golde, D. W., and Sherr, C. J. (1986). Evidence for the involvement of GM-CSF and FMS in the deletion (5q) in myeloid disorders. Science 231: 984-987. LeBeau, M. M., Epstein, N. D., O'Brien, S. J., Nienhuis, A. W., Yang, Y-C., Clark, S. C., and Rowley, J. D. (1987). The interleukin 3 gene is located on human chromosome 5 and is deleted in myeloid leukemias with a deletion of 5q. Proc. Natl. Acad. Sci. USA 84: 5913-5917. Nesbitt, M. N., and Francke, U. (1973). A system of nomenclature for band patterns of mouse chromosomes. Chromosoma 4 1 : 145-158. Schneider, C., Kingl R. M., and Phillipson, L. (1988). Genes specifically expressed at growth arrest of mammalian cells. Cell 54: 787-793. Solomon, E., Voss, R., Hall, V., Bodmer, W. F., Jass, J. R., Jeffreys, A. J., Lucibello, F. C., Patel, I., and Rider, S. H. (1987). Chromosome 5 allele loss in human colorectal carcinomas. Nature 328: 616-619. Wasmuth, J. J., Park, C., and Ferrell, R. E. (1989). Human Gene Mapping 10: Tenth International Workshop on Human Gene Mapping. Cytogenet. Cell. Genet. 51: 137-148.
13.
14.
549
Webb, G. C., Lee, J. S., Campbell, H. D., and Young, I. G. (1989). The genes for interleukins 3 and 5 map to the same locus on mouse chromosome 11. Cytogenet. Cell. Genet. 50: 107-110. Webb, G. C., Campbell, H. D., Lee, J. S., and Young, I. G. (1990). Mapping the gene for murine T-cell growth factor, IL-2, to bands B-C on chromosome 3 and for the a chain of the IL-2 receptor, IL-2ra, to bands A2-A3 on Chromosome 2. Cytogenet. Cell. Genet. 54: 164-168.
The Gene Encoding Human Plasma Carboxypeptidase B (CPB2) Resides on Chromosome 13 SIAO PING TSAI AND DENNIS DRAYNA1 Department of Molecular Biology, Genentech, Inc., 460 Pt. San Bruno Boulevard, South San Francisco, California 94080 Received February 25, 1992; revised July 13, 1992
H u m a n plasma carboxypeptidase B (CPB2) is a recently described carboxypeptidase, first isolated on the basis of its ability to bind plasminogen (4). Although the normal function of this protein is not well understood, it has several characteristics t h a t suggest t h a t it plays a role quite different from t h a t of the known pancreatic carboxypeptidase B (CPB1) (1). We undertook a study to map the C P B 2 gene and to compare its chromosomal location to t h a t of other carboxypeptidases. We used N I G M S hybrid cell mapping panel 2 in our study (3). This mapping panel contains 24 hybrid cell lines. In most cases these lines contain a single h u m a n chromosome on the rodent background, which is either mouse or hamster. We used P C R to determine the presence of the plasma carboxypeptidase B gene in the ~cell lines. We used P C R primers with the following sequences: primer P B P . P C R . 1 3 5' A T G AAG C T T T G C AGC C T T GCA 3' primer P B P . P C R . 1 4 5' CGC GAA GAC A T G C T G C T C ACA 3'. Th ese primers amplify a sequence within the region encoding the hydrophobic leader peptide of CPB2, and the expected P C R product is 63 bp in length. T h e CPB2-specific sequence was amplified using 30 cycles of 94°C for 1 min, 56°C for 2 min, and 72°C for 1.5 min, in reactions containing 0.5 #g of genomic DNA. Th e resultant products were separated on a 6% polyacrylamide gel and visualized by staining with ethidium bromide. T h e results of the P C R reactions using the N I G M S panel 2 D N A s are shown in Fig. 1. Our P C R primers did not amplify 1 To whom correspondence should be addressed. GENOMICS 14, 549-550 (1992) 0888-7543/92 $5.00 Copyright © 1992 by AcademicPress, Inc. All rights of reproduction in any form reserved.
550
BRIEF REPORTS
FIG. 1. Human plasma carboxypeptidase B-specific PCR products. PCR reactions and analysis of the PCR products were performed as described above. Lanes are labeled as follows: HU, HA, and MS are the parental human, hamster, and mouse DNAs, respectively. The numbers indicate the human chromosomes present in each hybrid DNA.
any specific sequence in either mouse or hamster DNA. Only a single hybrid cell line, GM/NA10898, produced an authentic 63-bp CPB2 product. This line contains chromosome 13 as its sole h u m a n component., This result, which is in agreement with results obtained from other somatic cell hybrid mapping panels including N I G M S hybrid mapping panel 1 and a commercially available hybrid panel (2, 3; results not shown), clearly indicates t h a t the C P B 2 coding sequence lies on chromosome 13. Two other cell lines, G M / N A 1 1 4 1 8 and G M / NA06318B, produced a smaller P C R product, approximately 45 bp in length. W h e t h e r these products represent an artifact or amplification of a related sequence on other chromosomes is not known. T h e assignment of C P B 2 to chromosome 13 contrasts this gene with those of other carboxypeptidases. For example, the h u m a n pancreatic carboxypeptidase A gene (CPA1) is known to reside on chromosome 7 (6), while the dipeptidyl carboxypeptidase 1 (angiotensin converting enzyme) gene resides on chromosome 17 (5). These results suggest t h a t the members of the carboxypeptidase gene family reside on many different chromosomes, analogous to those of the serine protease family (1).
REFERENCES 1.
Barrett, A. J., and MacDonald, J. K. (1986). "Mammalian Proteases, a Glossary and Bibliography," Vol. 2, pp. 155-224, Academic Press, London. 2. BIOS Corp., 291 Whitney Ave., New Haven, CT 06511. 3. Coriell Institute for Medical Research, 401 Haddon Ave., Camden, NJ 08103. 4. Eaton, D. L., Malloy, B. E., Tsai, S.-P., Henzel, W., and Drayna, D: (1991). Isolation, molecular cloning, and partial characterization of a novel carboxypeptidase B from human plasma. J. Biol. Chem. 266: 21833-21838. 5. Mattei, M. G., Hubert~ C., Alhenc-Gelas, F., Roeckel, N., Corvol, P., and Soubrier, F. (1989). The angioteusin-I converting enzyme gene is on chromosome 17. Cytogenet. Cell Genet. 51: 1041. 6. Stewart, E. A., Craik, C. S., Hake, L., and Bowcock, A. M. (1990). Human carboxypeptidase A identifies a BglII RFLP and maps to 7q31-qter. Am. J. Hum. Genes. 46: 795-800.
Assignment of the Human Gene Propionyl Coenzyme A Carboxylase, a-Chain, (PCCA) to Chromosome 13q32 by in Situ Hybridization Ingo Kennerknecht, 1 Christine Klett, and Horst Hameister Abtei/ung Klinische Genetik der Universit~t, Parkstrasse 1 I, W-7900 U/m, Germany
Received February 26, 1992
Propionic acidemia is an autosomal recessively inherited disorder of organic acid metabolism caused by deficiency of propionyl-CoA carboxylase [PCC, propanoyl-CoA: carbondioxide ligase (ADP-forming), EC 6.4.1.3]. The native enzyme is a large complex t h a t consists of two nonidentical subunits, most probably six a-subunits and six fl-subunits (6). c D N A clones coding for the a- and B-polypeptides of human PCC were isolated. By studying the segregation of D N A sequences homologous to the PCC cDNAs in a panel of m o u s e - h u m a n hybrids, the gene coding for the a-chain was mapped to chromosome 13 and t h a t for the B-chain was mapped to chromosome 3 (5). F u r t h e r refinement of localization of P C C A to 13q22--~q34 was possible by evaluation of a contiguous genetic linkage map of chromosome 13 (1). The P C C B gene was further regionalized to chromosome 3q13.3--~q22 by in situ hybridization (3). More recently, we have been able to tentatively assign the locus for the a-chain gene to 13q32 by gene dosage studies in cell lines with different chromosomal aberrations of chromosome 13 (2). We have now confirmed this assignment directly by use of a c D N A probe and in situ hybridization to metaphase chromosomes. For this purpose, chromosomes were prepared from P H A stimulated lymphocytes labeled with 20 ~g/pl bromo-desoxyuridine according to standard techniques. The recombinant a To whom correspondence should be addressed. GENOMICS 14, 550--551 (1992) 0888-7543/92 $5.00 Copyright © 1992 by AcademicPress, Inc. All rights of reproduction in any form reserved.