Cytogenet. Cell Genet. 22: 573-576 (1978)
Comparative gene mapping: the linkage relationships of the homologous genes for phosphoglucomutase and peptidase S are conserved in man and mouse P.A. L alley,1 U. F rancke,- and J.D. M inna1 ■NCI-VA Medical Oncology Branch, Veterans Administration Hospital, Washington, D.C., and -Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, Calif.
U.F. is supported by USPHS grant GM 21110 and by The National Foundation.
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Chinese hamster X mouse somatic cell hybrids, segregating mouse chromosomes, were employed to investigate the extent of conservation of autosomal linkage groups between mouse and man. During this study, the linkage relationships and chromosomal assignment of mouse peptidase S (PEP-S, E.C. 3.4.11.*) were investigated. Evidence is presented here which indicates that the gene coding for PEP-S is located on mouse chromosome 5, and the significance of this gene assignment for com parative gene mapping is discussed. Mouse and Chinese hamster PEP-S activity was separated using starch-gel electrophoresis (fig. 1). Chinese hamster (E36) PEP-S migrates as a single band of activity faster than mouse (RAG) PEP-S, which can be resolved into two bands of activity (fig. 1, channels 2 and 10). Hybrids which have lost mouse PEP-S show only the hamster band (channels 4 and 9), while in hybrids positive for mouse PEP-S, several heteropolymers were observed (at least four, possibly five) in addition to the parental bands (channels 3, 5, 6, 7 and 8). It is not clear whether both mouse bands are involved in heteropolymer formation, and, therefore, the number of true heteropolymers cannot be resolved. A similar pattern has been reported in human X Chinese hamster somatic cell hybrids.1 The segregation of PEP-S was compared to the segregation of 20 other enzyme markers in 37 Chinese hamster X mouse primary hybrid clones. A positive correlation was noted between PEP-S and PGM-1 (table I). Since the gene coding for PGM-1 is assigned to mouse chromosome 5,2 this result indicates that Pep-S can also be assigned to chromosome 5.
574
L alley et al.
Comparative mapping of gene loci *»
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The one discordant clone noted, which was PEP-S /PGM-1 +, was a result of chromosome breakage, since this clone contained no intact mouse chromosome but expressed several mouse enzymes. Chromosome and enzyme analysis of 20 hybrid clones demonstrated a perfect correlation between PEP-S expression and mouse chromosome 5 (table I). PEP-S segregated discordantly with all other chromosomes. Therefore, the gene coding for mouse PEP-S can be assigned to chromosome 5. Comparative mapping by homology may be acceptable for the X chro mosome.3 Obviously, such complete conservation is not seen for autosomes. However, the data presented in this report indicate that it may be possible to define autosomal regions which are conserved and thus suggest gene assignments by homology mapping for these regions. The genes coding for phosphoglucomutase-1 (PGM-1, homologous to human PGMs), /5-glucuronidase (/5GUS), malate dehydrogenase (mitochondrial) (MOR-1, homologous to human MDH ji), and PEP-S are located on mouse chro-
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Fig. 1. Mouse and Chinese hamster peptidase S was separated by starch-gel electro phoresis using a tris-EDTA-borate buffer system, pH 8.6, as described, using L-leucyl tyrosine as substrate.** Channels 1 and 2, mouse SC and RAG cells; 10, hamster E36; 3, 5, 6, 7, and 8, hybrids scored positive for mouse PEP-S; 4 and 9, hybrids scored negative for mouse PEP-S. ENS hybrids were formed from NZB mouse spleen cells X E36, and EBS hybrids were from BALB/c mouse spleen cells X E36 as described.!*
L alley et al.
Comparative mapping of gene loci
575
Table I. Segregation of peptidase-S (PEP-S) with mouse markers and chromosomes.“ Enzyme
DIP-1 ID-1 AK-1 PGD PGM-2 ENO-1 PGM-1 GPI LDH-A PEP-D APRT MOD-1 MPI PK-3 TRIP-1 ACP-1 ES-10 NP-1 DIP-1 HPRT
Chr
1 1 2 4 4 4 5 7 7 7 8 9 9 9 10 12 14 14 18 X
PEP-S/enzyime + /+
+ /-
-/+
16 11 16 13 12 13 20 19 19 18 17 16 16 16 14 17 16 15 12 17
5 7 4 8 9 8 0 2 2 2 3 5 5 5 7 4 5 5 9 1
8 7 8 4 4 4 1 9 9 8 4 3 4 4 8 9 5 5 4 14
Chr
8 6 8 12 12 12 10 7 7 6 12 13 12 12 8 7 11 9 11 0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 X Y
PEP-S/ch romosome + /+
+ /-
-/+
-/-
8 11 9 5 11 10 11 10 9 7 0 10 9 6 12 11 12 7 9 12 1
4 1
7 7 5 3 0 4 8 3 2 6 0 9 8 3 10 6 6 3 4 10 2
4 4 5 8 9 7 3 7 8 4 11 2 3 7 1 5 5 8 7 1 9
6 0 2 1 2 3 5 12 2 3 4 0 1 0 5 3 0 10
mosome 5.4 In man, the genes coding for /iGUS and MDH m are assigned to chromosome 7,5>c and PGM^> (homologous to mouse PGM-1) is assigned to chromosome 4.5 In the mouse, M or-l and Gus are 11 cM apart,1 while Pgm-1 and Gus are 36 cM apart.7 The regional localization of Pep-S in the mouse is not known. These data suggested that in man the gene coding for PEPS would be located on human chromosomes 4 or 7. It was found that human PEPS segregated concordantly with PGM> and could be assigned to chromosome 4 in man.1-8 Thus, the chromosome assignment of
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a Mouse and Chinese hamster enzymes were distinguished by starch-gel electro phoresis and specific enzyme staining as previously described.6 When available, mouse enzyme nomenclature is used; when not available, human nomenclature has been substituted. Mouse and Chinese hamster (E36) chromosomes were identified using Hoechst 33258 fluorescence and trypsin-Giemsa banding techniques.6 The chromosome assignments are as previously summarized.4,6
576
L alley et al.
Comparative mapping of gene loci
PEPS in man was suggested by homology mapping data from the mouse. As more genes within conserved linkage groups are identified, it may be possible to predict the assignment of genes already mapped in other mammalian species. We gratefully acknowledge the excellent assistance o f S. Stephenson , T. G re M. B rown , and W. Moss.
gorio ,
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1 B rown , S.; L alley, P.A., and M inna, J.D.: Assignment of the gene for pep tidase S (PEPS) to chromosome 4 in man and confirmation of peptidase D (PEPD) assignment to chromosome 19. This conference (1977). 2 Show s , T.B.; R uddle , F.H., and R oderick , T.H.: Phosphoglucomutasc electro phoretic variants in the mouse. Biochem. Genet. 3: 25-35 (1969). 3 O hno, S.: Ancient linkage groups and frozen accidents. Nature, Lond. 244: 259-262 (1973). 4 W omack, J.E.: Linkage map of the mouse. Mouse News Lett. 57: 6 (1977). 5 R uddle , F.H. and M eera K han, P.: Report of the committee on the genetic constitution of autosomes other than chromosomes 1, 2, and 6. Baltimore Conference (1975), pp. 31-53. 6 L alley, P.A.; B rown , J.A.; E ddy, R.L.; H aley, L.L.; B yers , M.G.; G oggin , A.P., and S how s , T.B.: Human /(-glucuronidase: assignment of the structural gene to chromosome 7 using somatic cell hybrids. Biochem. Genet. 15: 367-381 (1977). 7 L alley, P.A. and S how s , T.B.: Lysosomal and microsomal glucuronidase: genetic variant alters electrophoretic mobility of both hydrolases. Science 185: 442-444 (1974). 8 S how s , T.B.; Brow n , J.A.; Byers , M.G.; E ddy, R.L.; H aley, L.L., and G oggin , A.B.: Assignment of peptidase-S (PEPS) to chromosome 4 in man. This conference (1977). 9 F rancke, U.; L alley, P.A.; Moss, W.; I vy, J., and M inna, J.D.: Gene mapping in Mus musculus by interspecific cell hybridization: assignment of the genes for tripeptidase-1 to chromosome 10, dipeptidase-2 to chromosome 18, acid phosphatase-1 to chromosome 12, and adenylate kinase-1 to chromosome 2. Cytogenet. Cell Genet. 19: 57-84 (1977).
Comparative gene mapping: the linkage relationships of the homologous genes for phosphoglucomutase and peptidase S are conserved in man and mouse P.A. L alley,1 U. F rancke,- and J.D. M inna1 ■NCI-VA Medical Oncology Branch, Veterans Administration Hospital, Washington, D.C., and -Department of Pediatrics, University of California, San Diego, School of Medicine, La Jolla, Calif.
U.F. is supported by USPHS grant GM 21110 and by The National Foundation.
Downloaded by: Thomas Jefferson University Scott Library 147.140.27.100 - 8/24/2018 8:50:48 AM
Chinese hamster X mouse somatic cell hybrids, segregating mouse chromosomes, were employed to investigate the extent of conservation of autosomal linkage groups between mouse and man. During this study, the linkage relationships and chromosomal assignment of mouse peptidase S (PEP-S, E.C. 3.4.11.*) were investigated. Evidence is presented here which indicates that the gene coding for PEP-S is located on mouse chromosome 5, and the significance of this gene assignment for com parative gene mapping is discussed. Mouse and Chinese hamster PEP-S activity was separated using starch-gel electrophoresis (fig. 1). Chinese hamster (E36) PEP-S migrates as a single band of activity faster than mouse (RAG) PEP-S, which can be resolved into two bands of activity (fig. 1, channels 2 and 10). Hybrids which have lost mouse PEP-S show only the hamster band (channels 4 and 9), while in hybrids positive for mouse PEP-S, several heteropolymers were observed (at least four, possibly five) in addition to the parental bands (channels 3, 5, 6, 7 and 8). It is not clear whether both mouse bands are involved in heteropolymer formation, and, therefore, the number of true heteropolymers cannot be resolved. A similar pattern has been reported in human X Chinese hamster somatic cell hybrids.1 The segregation of PEP-S was compared to the segregation of 20 other enzyme markers in 37 Chinese hamster X mouse primary hybrid clones. A positive correlation was noted between PEP-S and PGM-1 (table I). Since the gene coding for PGM-1 is assigned to mouse chromosome 5,2 this result indicates that Pep-S can also be assigned to chromosome 5.
574
L alley et al.
Comparative mapping of gene loci *»
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The one discordant clone noted, which was PEP-S /PGM-1 +, was a result of chromosome breakage, since this clone contained no intact mouse chromosome but expressed several mouse enzymes. Chromosome and enzyme analysis of 20 hybrid clones demonstrated a perfect correlation between PEP-S expression and mouse chromosome 5 (table I). PEP-S segregated discordantly with all other chromosomes. Therefore, the gene coding for mouse PEP-S can be assigned to chromosome 5. Comparative mapping by homology may be acceptable for the X chro mosome.3 Obviously, such complete conservation is not seen for autosomes. However, the data presented in this report indicate that it may be possible to define autosomal regions which are conserved and thus suggest gene assignments by homology mapping for these regions. The genes coding for phosphoglucomutase-1 (PGM-1, homologous to human PGMs), /5-glucuronidase (/5GUS), malate dehydrogenase (mitochondrial) (MOR-1, homologous to human MDH ji), and PEP-S are located on mouse chro-
Downloaded by: Thomas Jefferson University Scott Library 147.140.27.100 - 8/24/2018 8:50:48 AM
Fig. 1. Mouse and Chinese hamster peptidase S was separated by starch-gel electro phoresis using a tris-EDTA-borate buffer system, pH 8.6, as described, using L-leucyl tyrosine as substrate.** Channels 1 and 2, mouse SC and RAG cells; 10, hamster E36; 3, 5, 6, 7, and 8, hybrids scored positive for mouse PEP-S; 4 and 9, hybrids scored negative for mouse PEP-S. ENS hybrids were formed from NZB mouse spleen cells X E36, and EBS hybrids were from BALB/c mouse spleen cells X E36 as described.!*
L alley et al.
Comparative mapping of gene loci
575
Table I. Segregation of peptidase-S (PEP-S) with mouse markers and chromosomes.“ Enzyme
DIP-1 ID-1 AK-1 PGD PGM-2 ENO-1 PGM-1 GPI LDH-A PEP-D APRT MOD-1 MPI PK-3 TRIP-1 ACP-1 ES-10 NP-1 DIP-1 HPRT
Chr
1 1 2 4 4 4 5 7 7 7 8 9 9 9 10 12 14 14 18 X
PEP-S/enzyime + /+
+ /-
-/+
16 11 16 13 12 13 20 19 19 18 17 16 16 16 14 17 16 15 12 17
5 7 4 8 9 8 0 2 2 2 3 5 5 5 7 4 5 5 9 1
8 7 8 4 4 4 1 9 9 8 4 3 4 4 8 9 5 5 4 14
Chr
8 6 8 12 12 12 10 7 7 6 12 13 12 12 8 7 11 9 11 0
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 X Y
PEP-S/ch romosome + /+
+ /-
-/+
-/-
8 11 9 5 11 10 11 10 9 7 0 10 9 6 12 11 12 7 9 12 1
4 1
7 7 5 3 0 4 8 3 2 6 0 9 8 3 10 6 6 3 4 10 2
4 4 5 8 9 7 3 7 8 4 11 2 3 7 1 5 5 8 7 1 9
6 0 2 1 2 3 5 12 2 3 4 0 1 0 5 3 0 10
mosome 5.4 In man, the genes coding for /iGUS and MDH m are assigned to chromosome 7,5>c and PGM^> (homologous to mouse PGM-1) is assigned to chromosome 4.5 In the mouse, M or-l and Gus are 11 cM apart,1 while Pgm-1 and Gus are 36 cM apart.7 The regional localization of Pep-S in the mouse is not known. These data suggested that in man the gene coding for PEPS would be located on human chromosomes 4 or 7. It was found that human PEPS segregated concordantly with PGM> and could be assigned to chromosome 4 in man.1-8 Thus, the chromosome assignment of
Downloaded by: Thomas Jefferson University Scott Library 147.140.27.100 - 8/24/2018 8:50:48 AM
a Mouse and Chinese hamster enzymes were distinguished by starch-gel electro phoresis and specific enzyme staining as previously described.6 When available, mouse enzyme nomenclature is used; when not available, human nomenclature has been substituted. Mouse and Chinese hamster (E36) chromosomes were identified using Hoechst 33258 fluorescence and trypsin-Giemsa banding techniques.6 The chromosome assignments are as previously summarized.4,6
576
L alley et al.
Comparative mapping of gene loci
PEPS in man was suggested by homology mapping data from the mouse. As more genes within conserved linkage groups are identified, it may be possible to predict the assignment of genes already mapped in other mammalian species. We gratefully acknowledge the excellent assistance o f S. Stephenson , T. G re M. B rown , and W. Moss.
gorio ,
Downloaded by: Thomas Jefferson University Scott Library 147.140.27.100 - 8/24/2018 8:50:48 AM
1 B rown , S.; L alley, P.A., and M inna, J.D.: Assignment of the gene for pep tidase S (PEPS) to chromosome 4 in man and confirmation of peptidase D (PEPD) assignment to chromosome 19. This conference (1977). 2 Show s , T.B.; R uddle , F.H., and R oderick , T.H.: Phosphoglucomutasc electro phoretic variants in the mouse. Biochem. Genet. 3: 25-35 (1969). 3 O hno, S.: Ancient linkage groups and frozen accidents. Nature, Lond. 244: 259-262 (1973). 4 W omack, J.E.: Linkage map of the mouse. Mouse News Lett. 57: 6 (1977). 5 R uddle , F.H. and M eera K han, P.: Report of the committee on the genetic constitution of autosomes other than chromosomes 1, 2, and 6. Baltimore Conference (1975), pp. 31-53. 6 L alley, P.A.; B rown , J.A.; E ddy, R.L.; H aley, L.L.; B yers , M.G.; G oggin , A.P., and S how s , T.B.: Human /(-glucuronidase: assignment of the structural gene to chromosome 7 using somatic cell hybrids. Biochem. Genet. 15: 367-381 (1977). 7 L alley, P.A. and S how s , T.B.: Lysosomal and microsomal glucuronidase: genetic variant alters electrophoretic mobility of both hydrolases. Science 185: 442-444 (1974). 8 S how s , T.B.; Brow n , J.A.; Byers , M.G.; E ddy, R.L.; H aley, L.L., and G oggin , A.B.: Assignment of peptidase-S (PEPS) to chromosome 4 in man. This conference (1977). 9 F rancke, U.; L alley, P.A.; Moss, W.; I vy, J., and M inna, J.D.: Gene mapping in Mus musculus by interspecific cell hybridization: assignment of the genes for tripeptidase-1 to chromosome 10, dipeptidase-2 to chromosome 18, acid phosphatase-1 to chromosome 12, and adenylate kinase-1 to chromosome 2. Cytogenet. Cell Genet. 19: 57-84 (1977).
