Enzymatic and metabolic studies on retrograde regulation mutants of yeast.
Two nuclear genes, RTG1 and RTG2, which sense the functional state of yeast mitochondria, have been described recently. Yeast strains with null alleles of either of these two genes (delta rtg1, delta rtg2) cannot grow on acetate as the sole carbon source and are auxotrophic for glutamate and aspartate. We report here a series of metabolic experiments and enzyme activity measurements that were made in an attempt to determine the reason for the acetate- phenotype and the glutamate/aspartate auxotrophy. Decreases in the activities (approximately 50%) in mitochondrial citrate synthase (CS1), acetyl-CoA synthetase, NAD isocitrate dehydrogenase, and pyruvate carboxylase were noted. When CS1 was overexpressed in the delta rtg1 and delta rtg2 mutants, these strains could grow on acetate but were still auxotrophic for glutamate/aspartate. We propose that, in the mutant strain, CS1 activity becomes limiting for efficient acetate utilization, but that other complex metabolic interactions are affected, limiting production of intermediates that would allow synthesis of glutamic and aspartic acids.
Small
W C
WC
Department of Veterans Affairs Medical Center, Dallas, Texas 75216, USA.
Brodeur
R D
RD
Sandor
A
A
Fedorova
N
N
Li
G
G
Butow
R A
RA
Srere
P A
PA
D000106:Acetate-CoA Ligase / Q000378:metabolism; D001224:Aspartic Acid / Q000378:metabolism; D002950:Citrate (si)-Synthase / Q000096:biosynthesis / Q000378:metabolism; D005649:Fumarate Hydratase / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D018698:Glutamic Acid / Q000378:metabolism; D007521:Isocitrate Dehydrogenase / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D016296:Mutagenesis; D010101:Oxygen Consumption; D011766:Pyruvate Carboxylase / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013045:Species Specificity; D013385:Succinate Dehydrogenase / Q000378:metabolism
D001224:Aspartic Acid; D018698:Glutamic Acid; D008291:Malate Dehydrogenase; D007521:Isocitrate Dehydrogenase; D013385:Succinate Dehydrogenase; D002950:Citrate (si)-Synthase; D005649:Fumarate Hydratase; D000106:Acetate-CoA Ligase; D011766:Pyruvate Carboxylase