Biochemical and molecular analysis of spontaneous and induced mutations at the mouse Mod-1 locus.
We have analyzed five Mod-1 (malic enzyme) mutants at the molecular and biochemical level. Four of these mutants, three electrophoretic variants and one null mutant, were induced by ethylnitrosourea (ENU). Another null mutant was the result of a spontaneous mutation. All of these mutations were heritable in a Mendelian fashion and viable in the homozygous condition. Restriction endonuclease and Southern blot analysis revealed that the spontaneous null mutant possessed an altered restriction fragment banding pattern. All of the ENU-induced mutants possessed normal restriction fragment banding patterns. All 5 mutants produced normal levels of Mod-1-specific mRNA. Only the spontaneous null mutant produced mRNA with altered size, which was consistent with the altered DNA-banding pattern. MOD-1 enzyme activity levels were normal in the three ENU-induced mutants with altered electrophoretic mobility. Enzyme activity was significantly lower than normal in tissues from animals homozygous for the null alleles, however, using Western blot analysis, low but significant levels of MOD-1 protein in Mod-1 null homozygotes were detected.
Cobb
R R
RR
Chemistry and Life Sciences Group, Research Triangle Institute, Research Triangle Park, NC 27709.
Burkhart
J G
JG
Dubins
J S
JS
Barnett
L B
LB
Lewis
S E
SE
D000818:Animals; D003433:Crosses, Genetic; D004262:DNA Restriction Enzymes; D004592:Electrophoresis, Starch Gel; D005038:Ethylnitrosourea; D015870:Gene Expression; D005796:Genes; D015151:Immunoblotting; D008099:Liver / Q000201:enzymology; D008291:Malate Dehydrogenase / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D009154:Mutation; D012333:RNA, Messenger / Q000235:genetics
D012333:RNA, Messenger; D008291:Malate Dehydrogenase; D004262:DNA Restriction Enzymes; D005038:Ethylnitrosourea