Biochemical studies of pyrithiamine-resistant mutants of Escherichia coli K12.
The biochemical properties of strains (PT-R101 and PT-R108) of Escherichia coli K12 resistant to growth inhibition by pyrithiamine, and antimetabolite of thiamine, have been studied. Intracellular thiamine pyrophosphate concentration in these resistant strains was slightly but definitely higher than that in the parent strain. Thiamine synthesis from the pyrimidine and thiazole moieties of thiamine by cell suspensions was greater in the resistant strains than the parent strain. The activities of enzymes involved in thiamine biosynthesis in these pyrithiamine-resistant strains were 2-3 times higher than the parent strain (3301), except for thiamine-phosphate kinase, which was indetectable in in vitro assay of the activity. However, other evidence indicates that this enzyme is not defective but is functioning in vivo and, furthermore, that the negligible activity of this enzyme did not affect the growth rate of the mutants. The activities of these enzymes were further enhanced when PT-R101 was grown on 5mM adenine and were reduced almost to zero when the strain was grown on 0.1 muM thiamine in the same way as the parent strain. However, when these resistant strains were grown on a low concentration of thiamine such as 0,05 muM, thiamine synthesis by cell suspensions also decreased, but only to a limited extent compared with the parent strain. These results suggest that PT-R101 and PT-R108 are altered in the mechanisms of regulation of thiamine biosynthesis. Their altered properties might be due to a reduced binding affinity of the repressor protein, which is involved on the regulation of thiamine synthesis, for the corepressor, thiamine pyrophosphate,
D000225:Adenine / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D009154:Mutation; D011726:Pyridinium Compounds / Q000494:pharmacology*; D013045:Species Specificity; D013831:Thiamine / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000494:pharmacology; D013835:Thiamine Pyrophosphate / Q000378:metabolism
D011726:Pyridinium Compounds; D000225:Adenine; D013835:Thiamine Pyrophosphate; D013831:Thiamine