In vivo hydrolysis of S-adenosyl-L-methionine in Escherichia coli increases export of 5-methylthioribose.
Escherichia coli can not synthesize methionine from 5-methylthioribose (MTR) but instead exports this sulfur-containing, energy-rich molecule into the surrounding medium. Transforming E. coli with plasmids that direct expression of the cloned coliphage T3 S-adenosyl-L-methionine (SAM) hydrolase (SAMase) induces the met regulon by cleaving the SAM co-repressor to form 5'-methylthioadenosine, which is then cleaved to produce MTR. To test the effect of in vivo SAMase activity on MTR production and its fate, cultures were incubated in the presence of [35S]methionine and [methyl-3H]methionine. Cells with SAMase activity produced significantly enhanced levels (up to 40-fold in some trials) of extracellular MTR -- the only radiolabeled compound released in significant amounts -- when compared with controls. SAM synthetase (metK) mutants transformed with SAMase expression vectors did not show this increase, verifying the path through SAM as the sole route to MTR production. SAMase expression had little or no effect on intracellular MTR pools, levels of radiolabeled macromolecules, or the transfer of methyl groups to compounds that could be precipitated by trichloroacetic acid. Thus, MTR appears to be a dead-end metabolite in E. coli, begging questions about how this has evolved, the mechanism of MTR export for the cell, and whether the release of MTR is important for some other activity.
Hughes
Jeffrey A
JA
Department of Biology, Hanover College, IN 47243, USA. hughes@hanover.edu
D001692:Biological Transport; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism; D006868:Hydrolysis; D008715:Methionine / Q000493:pharmacokinetics; D010957:Plasmids / Q000235:genetics; D012436:S-Adenosylmethionine / Q000378:metabolism*; D013865:Thioglycosides / Q000378:metabolism*; D014169:Transformation, Bacterial
D013865:Thioglycosides; C001955:5-methylthioribose; D012436:S-Adenosylmethionine; D008715:Methionine; D006867:Hydrolases; C019614:adenosylmethionine hydrolase