Genetic control of two electrophoretic variants of glucosephosphate isomerase in the mouse (Mus musculus)

The autosomal variation and the genetic control of GPI has been determined by a comparison of electrophoretic patterns of F₁ and backcross progeny of three inbred strains of mice. The locus controlling the production of GPI in the mouse has been designated Gpi-1. Two alleles at this locus have been described and designated Gpi-1 ^a and Gpi-1 ^b, which represent, respectively, the slow and fast electrophoretic forms. Twenty-seven inbred strains of mice have been classified for these two alleles. The absence of close linkage of Gpi-1 to seven other genetic loci has been determined. It has been demonstrated that the polymorphism of Gpi-1 is widely distributed in feral mice. GPI was expressed in vitro and in four types of malignant tumors.Supported by U.S. Public Health Service Grants GM-09966, from General Medical Sciences, and GY 4193.     

A null mutation at the mouse Phosphoglucomutase-1 locus and a new locus,Pgm-3

A null mutation at the phosphoglucomutase locus (Pgm-1) was discovered by electrophoretic analysis of the inbred mouse strain C57 BL/6J. The null allele (Pgm-1 ⁿ) was shown to segregate as a Mendelian unit alternative to the Pgm-1 ^a and Pgm-1 ^b alleles. Mice expressing the Pgm-1 ⁿ allele, either in the heterozygous or homozygous state, are viable, healthy, and fertile. The occurrence of the Pgm-1 ⁿ mutant revealed a previously unreported genetic locus (Pgm-3) that controls the expression of a third phosphoglucomutase. Two electrophoretically expressed alleles of Pgm-3 (inherited without dominance) are found in the inbred mouse strains C57 BL/6J and DBA/2J. Linkage observed between the Pgm-3 locus, the dilute locus (d) and the cytoplasmic malic enzyme locus (Mod-1) has allowed assignment of the Pgm-3 locus to chromosome 9. A striking tissue specific expression of Pgm-1 and Pgm-3 was observed. Products of the Pgm-3 locus were detected in kidney, testes, brain, and heart. In contrast, Pgm-1 controlled isozymes were present in kidney, spleen, ovaries, and erythrocytes.Financial support for this work was provided in part by Contract #263-78-C-0393 from the National Institute of Environmental Health Sciences to the Research Triangle Institute.     

A genetically determined variant of the a-subunit of lactic dehydrogenase in the deer mouse

Analysis by starch gel electrophoresis of erythrocyte hemolysates and brain extracts indicated that an electrophoretic variant of lactic dehydrogenase discovered in Peromyscus involves the A subunit. The tetramers containing the variant molecules migrate more rapidly than the normal ones, and the staining reactions suggested a reduced enzymatic activity. Comparisons of the erythrocyte LDH activity of normal and homozygous variant animals indicated that the variant enzyme is intrinsically less active than the normal. The variant is inherited as an autosomal codominant and appears to confer no selective disadvantage.This work was supported in part by a grant GM-15885 from the National Institutes of Health, U.S. Public Health Service, and in part by a supporting fund established in the name of Zachary Pitts Research Fellowship.     

Phosphoglucomutase isozymes of red cells in Mus musculus: Additional polymorphism at PGM-1 compared by two electrophoretic systems

Phosphoglucomutase (PGM) of red cells was examined in 15 inbred strains of mice, using two different starch gel electrophoretic buffer systems. Two new alleles, Pgm-1 ^c and Pgm-1 ^d, were discovered at the Pgm-1 locus. Pgm-1 ^c was first identified in strain C3H/HeNWe and Pgm-1 ^d in 129/ReWl. No variation was observed at the Pgm-2 locus.Supported in part by USPHS Pre-doctoral Fellowship No. 5 F1 GM-32,680, USPHS Research Resources Grant No. 5-PO6 RR 00343-05, USPHS (National Cancer Institute) Chemotherapy Contract 71-2010, and Biomedical Sciences Support Grant FR 07037 to the University of Kansas.     

Electrophoretic variation in human serum ceruloplasmin: A new genetic polymorphism

Through the application of a specific oxidase stain to results of starch gel electrophoresis of human serum, three different electrophoretic forms of ceruloplasmin—denoted CpA (fast), CpB (intermediate), and CpC (slow)—have been defined. The electrophoretic differences are small and were first recognized through a rare variant individual who had only the fast and slow forms. Five phenotypes displaying different combinations of the three electrophoretic forms have been defined in American Negroes; these are called CpA, CpAB, CpB, CpAC, and CpBC. Twin, family, and population studies have yielded evidence indicating that the A and B electrophoretic forms are controlled by a pair of autosomal codominant alleles, designated Cp ^A and Cp ^B , and suggesting that the C form may be determined by a third allele, Cp ^C , at the same locus. The variants constitute a genetic polymorphism in American Negroes, but occur only rarely in Caucasians.Supported by U.S. Atomic Energy Commission Contract AT(11-1)-1552, by U.S. Public Health Service Research Grants AM 09381 and HD 02083, and by U.S. Public Health Service Career Development Awards 6-K3-HE-24, 980 (DCS) and 1-K3-A-7959 (GJB).     

Genetic variation in the carbonic anhydrase isozymes of macaque monkeys. II. Inheritance of red cell carbonic anhydrase levels in different carbonic anhydrase I genotypes of the pig-tailed macaque,Macaca nemestrina

The inheritance of red blood cell levels of carbonic anhydrase isozymes (CA I and CA II) has been studied in different carbonic anhydrase I genotypes of the pig-tailed macaque, Macaca nemestrina. Quantitation of CA I isozymes in a series of animals indicates that the total CA I concentration is the sum of the average effects of each CA I structural allele and that the average effects are independent of the various allelic combinations. The relative average effects were 0.32:0.95:1.0 for the CA I ^a, CA I^b, and CA I ^c structural genes, respectively. It is also demonstrated that the level of CA II is related to the CA I genotypes. Multiple regression analysis demonstrated that each dose of CA I-deficiency gene present decreased the CA II concentration by approximately 30%, with this decrease in CA II level being solely related to the dose of CA I-deficiency gene and not to the level of CA I. The CA I-deficient animals produce CA I products that are similar to the common CA Ia, CA Ib, CA Ic electrophoretic types. Limited mating data indicate that the CA I components in CA I-deficient animals are inherited codominantly.Supported by U.S. Public Health Service Research Grant GM-15419.This report is a portion of a dissertation submitted to the University of Michigan in partial fulfillment of the requirements for the Doctor of Philosophy degree.U.S. Public Health Service Predoctoral Trainee (GM-71-14).     

Linkage of isozyme loci in the mouse: Phosphoglucomutase-2 (Pgm-2), mitochondrial NADP malate dehydrogenase (Mod-2), and dipeptidase-1 (Dip-1)

The linkages of the isozyme genes Mod-2, Pgm-2, and Dip-1 have been determined in tests with established linkage group markers among inbred strains of mice. Unique alleles for both Mod-2 and Pgm-2 have been observed in the strain of SM/J. Linkage was determined from backcross progeny of the matings C57BL/6J×(SM/J×C57BL/6J)F₁, (SM/J×SWR/J)F₁×SM/J, and (SM/J×SWR/J)F₁×SJL/J. The gene Mod-2 is on linkage group 1. In a three-point cross of the loci Gpi-1, c, and Mod-2, the c locus was determined to be the middle gene. No double crossovers were observed. Our combined data show the following linkages: Gpi-1 to c, 28.3±3.2%; Gpi-1 to Mod-2, 33.3±3.0%; and c to Mod-2, 4.1±2.8%. The proposed gene order for four markers on LG I is Gpi-1-p-c-Mod-2. The gene Pgm-2 was linked to Gpd-1 (27.0±4.2%) on LGVIII. Two backcrosses segregating for Pgm-2 and b, (SM/J×DBA/2J) F₁×DBA/2J and (SM/J×DBA/2J)F₁×C57BR/cdJ, showed 9.1±4.3% recombination. The proposed gene order on LG VIII is b-Pgm-2-Gpd-1. The genes Pgm-1 and Pgm-2 are not linked (53.4±4.4%). Linkage of the isozyme genes Dip-1 and Id-1 on LG XIII was observed in backcross progeny of the crosses (SJL/J×C57BL/6J)F₁×SJL/J and C57BL/6J×(SM/J×C57BL/6J)F₁. The combined recombination was 23.8±2.8%. Two cases are established where genes whose enzyme products share substrate affinities (Pgm-1 and Pgm-2; Mod-1 and Mod-2) are not linked. Our data generally support the conclusion that functionally or metabolically related isozyme genes are not contiguous on mouse linkage groups.This investigation was supported in part by Public Health Service General Research Support Grant GM-09966 and in part by Public Health Service Training Grant 5T01 HD-00032-07 from the National Institute of Child Health and Human Development, and by Atomic Energy Commission contract AT(30-1)-3671.     

Genetic variation in the carbonic anhydrase isozymes of macaque monkeys. I. The radioimmunosorbent assay

A radioimmunosorbent technique is described which is capable of independently detecting both isozymes of carbonic anhydrase, CA I and CA II, in concentrations as low as 1 ng/ml. The technique is used to quantitate the different electrophoretic variants of red cell CA I as well as levels of CA II in the pig-tailed macaque, Macaca nemestrina.Supported by U.S. Public Health Service research grant GM-15419.U.S. Public Health Service Predoctoral Trainee (GM-71-14).     

Polymorphism and linkage for mannosephosphate isomerase in Mus musculus

Electrophoretic variants of the enzyme mannosephosphate isomerase (E.C. 5.3.1.8) (MPI) have been discovered in the mouse. The MPI-IA phenotype was found in Mus castaneus and in the inbred strain MA/J of Mus musculus. Other inbred strains of Mus musculus examined possessed the MPI-1B phenotype. Genetic studies show that the MPI variants segregate as codominant alleles of a single autosomal locus, designated Mpi-1 in linkage group II. The gene order Mod-1-d-Mpi-1 has been established. Human and murine forms of MPI differ and can be detected in in vitro fibroblasts and somatic cell hybrid populations.Supported by GB 18543 of the National Science Foundation, RSA-70-20 of the Connecticut Research Commission, and GM 09966 of the U.S. Public Health Service.     

Linkage of genes for laminin B1 and B2 subunits on chromosome 1 in mouse

Summary We have used cDNA clones for the B1 and B2 subunits of laminin to find restriction fragment length DNA polymorphisms for the genes encoding these polypeptides in the mouse. Three alleles were found forLamB2 and two forLamB1 among the inbred mouse strains. The segregation of these polymorphisms among recombinant inbred strains showed that these genes are tightly linked in the central region of mouse Chromosome 1 betweenSas-1 andLy-m22, 7.4±3.2 cM distal to thePep-3 locus. There is no evidence in the mouse for pseudogenes for these proteins. This work was supported by U. S. Public Health Service Grant GM28464 to R.W.E. Editor's Statement Investigation into the biosynthesis of laminin indicates that several different polypeptides are assembled to form the intact molecule. This paper represents an extension of previous work which takes a genetic approach to further define the relationships among the polypeptides involved. Gordon H. Sato     

Chemical characterization of a new Japanese variant of carbonic anhydrase I,CA INagasaki 1 (76 Arg→Gln)

A new inherited variant of carbonic anhydrase I (CA I), designated CA I_{Nagasaki 1} (CA I_{NGS 1}), was discovered during a survey of hemolysates from 5852 individuals from the cities of Hiroshima and Nagasaki in Japan. Analysis of the amino acid composition of a tryptic peptide from the CA I_{NGS 1} variant indicated that a glutaminyl residue was substituted for an arginyl residue at position 76. Heat degradation studies showed that the CA I_{NGS 1} variant was less stable than normal CA I. The CO₂ hydrase and esterase activities of the normal and variant carbonic anhydrases I, as well as the relative amounts of the two enzymes in heterozygotes, were similar.This work was supported in part by Contract E(11-1)-1552 with the Energy Research and Development Administration, Washington, D.C. (to J. V. Neel), and by U.S. Public Health Service Grant GM-24681.     

Duplication of the autosomally inherited 6-phosphogluconate dehydrogenase gene locus in tetraploid species of cyprinid fish

Among members of the fish family Cyprinidae,a diploid—tetraploid relationship exists. The present study on electrophoretic patterns of 6-phosphogluconate dehydrogenase indicates that such diploid members as Barbus tetrazonamaintain allelic polymorphism at a single gene locus for this enzyme. Tetraploid members such as the carp and goldfish are endowed with two separate gene loci for 6-PGD. Tetraploid evolution apparently fixed two former alleles of the same locus as two separate gene loci. Furthermore, it appears that after becoming tetraploid, the carp and goldfish developed a separate regulatory mechanism for each locus; thus preferential activation of one or the other 6-PGD locus occurs in different tissues of tetraploid species. This investigation was supported in part by a grant (CA-05138) from the National Cancer Institute, U.S. Public Health Service, and in part by a research fund established in honor of General James H. Doolittle. Contribution No. 4-68, Department of Biology, City of Hope Medical Center.Dr. Bender is a recipient of International Postdoctoral Fellowship 3 F05-TW-01198-0152 from the U.S. Public Health Service.     

Linkage of Pgm-3 in the house mouse and homologies of three phosphoglucomutase loci in mouse and man

The discovery of a third phosphoglucomutase locus (Pgm-3) in the house mouse is reported. Three alleles are recognized on the basis of differences in electrophoretic mobility and enzymatic activity. Pgm-3 ^a (fast mobility and high activity) is present in inbred strain C57BL/10J and 24 other strains; Pgm-3 ^b (slow mobility and high activity) is present in LP/Pas and six other strains; and Pgm-3 ^c (no detectable activity in any tissue tested) is present in strain DBA/2J and 14 other strains. Seventy-four recombinant inbred strains derived from progenitors that differed at Pgm-3 were used to study genic linkage. Pgm-3 is on chromosome 9 and is linked to Sep-1, d, Mod-1, and Ltw-3. Gene order and recombination frequencies are estimated as d 3.8±1.8% Pgm-3 2.3±1.2% Mod-1. Substrate specificities and cofactor requirements show that mouse Pgm-1 is homologous with human Pgm-2, mouse Pgm-2 with human Pgm-1, and mouse Pgm-3 with human Pgm-3.This research was supported in part by NIH Research Grant GM18684 from the National Institute of General Medical Sciences to B.A.T. and by grants from NIH A105531-02 and the Volkswagon Foundation to Jan Klein. J.H.N. was a recipient of a Fellowship from the Max-Planck-Gesellschaft, Munich. G.S. and J.K. were supported by funds from the Deutsche Forschungsgemeinschaft.     

Genetics of glucose phosphate isomerase and phosphoglucomutase in Aedes albopictus (diptera: culicidae)

Summary Glucose phosphate isomerase (E.C. 5.3.1.9) and phosphoglucomutase (E.C. 2.7.5.1) were found to be polymorphic in a laboratory colony of Aedes albopictus. The glucose phosphate isomerase locus is represented by two alleles resulting in three genotypes, while the phosphoglucomutase locus is represented by at least five alleles giving rise to a total of 15 genotypes. The inheritance of these two enzymes is of the Mendelian type with codominant alleles. Present data indicate that these genes are not linked.Of 105 mosquitoes analysed for these two gene-enzyme systems, the frequencies for glucose phosphate isomerase alleles are Gpi ^S=0.68 and Gpi ^F=0.32, while the frequencies for phosphoglucomutase alleles are Pgm ^A=0.16, Pgm ^B=0.11, Pgm ^C=0.19, Pgm ^D=0.30 and Pgm ^F= 0.24. The frequencies of the three glucose phosphate isomerase genotypes are in accord with Hardy-Weinberg expectations (X ₁ ² =2.74). Similarly, the frequencies of the 15 phosphoglucomutase genotypes probably do not differ significantly from Hardy-Weinberg expectations (X ₁₀ ² = 18.45).     

The amino acid substitution and some chemical properties of a variant human erythrocyte carbonic anhydrase: Carbonic anhydrase IdMichigan

Carbonic anhydrase Id_Michigan, an electrophoretic variant of human red cell carbonic anhydrase I, was purified from erythrocytes obtained from an individual heterozygous for the trait. Primary structural analysis indicates that a lysine residue has exchanged for a threonine residue in the variant enzyme. After isolation, there was approximately 1.8 times as much normal as variant enzyme. Thermostability studies demonstrated that carbonic anhydrase Id was more thermolabile than the normal enzyme. The normal and variant enzymes showed no differences in specific carboxylesterase activity or CO₂ hydratase activity. Utilizing the carboxylesterase activity toward -naphthyl acetate, the normal and variant enzymes had similar Michaelis constants, pH profiles, and rates of inhibition by acetazolamide. Immunochemical studies did not demonstrate an antigenic difference for the variant enzyme.Supported in part by Research Grants 2 T1 GM-76, 5 TO1 GM 00071-09, and GM 09252 from U.S. Public Health Service.This report is a portion of a dissertation submitted to the University of Michigan in partial fulfillment of the requirements for the doctor of philosophy degree.     

Electrophoretic and heat-stability polymorphism at the phosphoglucomutase (Pgm) locus in natural populations of Drosophila melanogaster

Genetic polymorphism for electrophoretic and heat-sensitive alleles is known at the phosphoglucomutase (Pgm) locus in Drosophila melanogaster. Analysis of the distribution of electrophoretic and thermosensitive (ts) alleles was carried out in natural populations from Canada and West Africa and compared with already known data on Italian populations [Trippa, G., Loverre, A., and Catamo, A. (1976). Nature 260:42]. The data show the existence of five common alleles, Pgm ^1.00,tr, Pgm ^1,00,ts, Pgm ^0.70,ts, Pgm ^1.20,ts, and Pgm ^1.50,tr, and two rare alleles, Pgm ^0.55,ts and Pgm ^1.20,tr. The most frequent allele is always Pgm ^1.00,tr; the second most common allele is always of the ts type. The cumulated frequencies of ts alleles in the populations varies between 11 and 32%. The heat stability polymorphism is present in all populations examined and shows again the uniform geographic pattern that has been found for electrophoretic variation at this locus.This research was partially supported by an operating grant (to G.R.C.) from the Canadian National Science and Engineering Research Council (NSERC).     

Mitochondrial malate dehydrogenase and malic enzyme: Mendelian inherited electrophoretic variants in the mouse

Malate dehydrogenase and malic enzyme each possess supernatant and mitochondrial molecular forms which are structurally and genetically independent. We describe electrophoretic variants of the mitochondrial enzymes of malate dehydrogenase and malic enzyme in mice. Progeny testing from genetic crosses indicated that the genes which code for mitochondrial malate dehydrogenase and malic enzyme were not inherited maternally but as independent unlinked nuclear autosomal genes. The locus for mitochondrial malic enzyme was located on linkage group I. Linkage analysis with a third mitochondrial enzyme marker, glutamic oxaloacetic transaminase, showed that the nuclear genes which code for the three mitochondrial enzymes were not closely linked to each other. This evidence suggests that clusters of nuclear genes coding for mitochondrial function are unlikely in mice.Supported by U.S. Public Health Service grants 5F2 HD-35,531 and GM-09966.     

The predominance of heterozygotes found in wild goldfish of lake Erie at the gene locus for sorbitol dehydrogenase

Sorbitol dehydrogenase (SDH) found in liver, kidney, and gonads of the goldfish (Carassius auratus) appears to be specified by a single autosomally inherited gene locus and, when subjected to electrophoresis, SDH behaves as a tetramer in that a homozygote gives a single band, while a heterozygote shows five bands of SDH. In a population of the wild goldfish inhabiting Lake Erie, two alleles which specify electrophoretic variants S^A and S^B coexist in nearly equal frequency, and there appears to be a slight excess of heterozygotes. Sixty-seven of the 109 wild goldfish were S^A/S^B heterozygotes. In the domesticated goldfish, on the other hand, an allele for the S^B variant appears to be a very rare allele. Only one of the 100 domesticated goldfish studied here and none of the 100 domesticated goldfish studied in Germany was an S^A/S^B heterozygote.Supported in part by a grant (CA 05138) and grant (FR 00433) from the National Cancer Institute, U.S. Public Health Service.On leave from McMaster University, Hamilton, Ontario, Canada.     

Genetic polymorphism of the sixth component of complement (C6) in mice

Immunofixation after isoelectric focusing revealed two forms of mouse C6, C6A and C6M, both of which consist of two major protein bands and one or more acidic minor bands. They were distinguishable by their different isoelectric point (pI) ranges: C6M has more acidic pI ranges (pH < 6.2) than C6A (pH < 6.3). C6A was found in common inbred mice of Mus musculus domesticus, while C6M was found in inbred and wild mice of M. m. molossinus (Japanese wild mice, an Asian subspecies). Breeding experiments showed that these two forms of C6 were controlled by a single codominant autosomal locus. We propose the designation C-6 for this locus with two alleles, C-6 ^a and C-6 ^m , which encode for C6A and C6M, respectively. Linkage analysis indicated that the locus is not closely linked to the following loci: Idh-1, agouti, Amy-1, brown, Gpd-1, Mup-1, Pgm-2, Pgm-1, albino, Hbb, Es-1, Mod-1, Sep-1, Es-3, Igh-1, beige, Es-10, Sod-1, and C-3.     

Metabolic suppressors of a regulatory mutant in Neurospora

The synthesis of a number of enzymes of sulfur assimilation in Neurospora crassa is controlled by the sulfur source. Mutants in one regulatory gene, cys-3, are unable to make any of the enzymes. This locus is thought to specify a macromolecule that is required for the expression of the structural genes. A mutant, scon ^c, of another regulatory gene is nonrepressible for the synthesis of the enzymes. We report here the isolation of suppressed scon ^c strains which are actually the double mutant, scon ^c,cys-3. These strains are phenotypically indistinguishable from single mutants at the cys-3 locus. Thus cys-3 is epistatic to scon ^c. Evidence that the expression of the cys-3 gene is itself controlled is also presented.This work is supported by a Public Health Service grant, GM-08995, from the National Institute of General Medical Sciences. One of us (R.L.M.) is supported by a Public Health Service Career Development Award.