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PMID:6178725

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Citation

Ryals, J, Hsu, RY, Lipsett, MN and Bremer, H (1982) Isolation of single-site Escherichia coli mutants deficient in thiamine and 4-thiouridine syntheses: identification of a nuvC mutant. J. Bacteriol. 151:899-904

Abstract

A method is described to rapidly select and classify many independent near-UV irradiation-resistant Escherichia coli mutants, which include tRNA modification and RNA synthesis control mutants. One class of these mutants was found to be simultaneously deficient in thiamine biosynthesis and in the ability to modify uridine in tRNA to 4-thiouridine, known to be the target for near-UV irradiation. These mutants were found to be unable to make thiazole, a thiamine precursor. The addition of thiazole restores the thiamine deficiency but does not render the cells near-UV irradiation sensitive. In vitro studies on one of these mutants indicated a deficiency in protein factor C (nuvC), required for the 4-thiouridine modification of tRNA. In P1 transduction, the thiazole marker cotransduced with the histidine marker, which places the thiazole marker between 42 and 46 min on the E. coli chromosome map. Both thiamine production and 4-thiouridine production were resumed by 87% of the spontaneous reversions, suggesting a single-point mutation. Our results indicate that we have isolated nuvC mutants and that the nuvC polypeptide is involved in two functions, tRNA modification and thiazole biosynthesis.

Links

PubMed PMC220340

Keywords

Chromosome Mapping; Chromosomes, Bacterial; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli/radiation effects; Genes, Bacterial; Mutation; RNA, Bacterial/metabolism; RNA, Transfer/metabolism; Thiamine/biosynthesis; Thiazoles/biosynthesis; Thiouridine/biosynthesis; Ultraviolet Rays

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ECOLI:THII

involved_in

GO:0002937: tRNA 4-thiouridine biosynthesis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:THII

involved_in

GO:0009228: thiamine biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

Notes

See also

References

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