Thioredoxin or glutaredoxin in Escherichia coli is essential for sulfate reduction but not for deoxyribonucleotide synthesis.
We have shown previously that Escherichia coli cells constructed to lack both thioredoxin and glutaredoxin are not viable unless they also acquire an additional mutation, which we called X. Here we show that X is a cysA mutation. Our data suggest that the inviability of a trxA grx double mutant is due to the accumulation of 3'-phosphoadenosine 5'-phosphosulfate (PAPS), an intermediate in the sulfate assimilation pathway. The presence of excess cystine at a concentration sufficient to repress the sulfate assimilation pathway obviates the need for an X mutation and prevents the lethality of a novel cys+ trxA grx double mutant designated strain A522. Mutations in genes required for PAPS synthesis (cysA or cysC) protect cells from the otherwise lethal effect of elimination of both thioredoxin and glutaredoxin even in the absence of excess cystine. Both thioredoxin and glutaredoxin have been shown to be hydrogen donors for PAPS reductase (cysH) in vitro (M. L.-S. Tsang, J. Bacteriol. 146:1059-1066, 1981), and one or the other of these compounds is presumably essential in vivo for growth on minimal medium containing sulfate as the sulfur source. The cells which lack both thioredoxin and glutaredoxin require cystine or glutathione for growth on minimal medium but maintain an active ribonucleotide reduction system. Thus, E. coli must contain a third hydrogen donor active with ribonucleotide reductase.
Russel
M
M
Laboratory of Genetics, Rockefeller University, New York, New York 10021.
Model
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Holmgren
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A
D000227:Adenine Nucleotides / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000378:metabolism*; D003553:Cystine / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D005838:Genotype; D054477:Glutaredoxins; D009154:Mutation; D010084:Oxidation-Reduction; D010088:Oxidoreductases; D010724:Phosphoadenosine Phosphosulfate / Q000096:biosynthesis*; D011506:Proteins / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism*
D000227:Adenine Nucleotides; D001426:Bacterial Proteins; D003854:Deoxyribonucleotides; D054477:Glutaredoxins; D011506:Proteins; D013431:Sulfates; D010724:Phosphoadenosine Phosphosulfate; D003553:Cystine; D013879:Thioredoxins; D010088:Oxidoreductases