In vivo hydrolysis of S-adenosylmethionine induces the met regulon of Escherichia coli.
Regulation of methionine biosynthesis in Escherichia coli involves a complex of the MetJ aporepressor protein and S-adenosylmethionine (SAM) repressing expression of most genes in the met regulon. To test the role of SAM in the regulation of met genes directly, SAM pools were depleted by the in vivo expression of the cloned plasmid vector-based coliphage T3 SAM hydrolase (SAMase) gene. Cultures with in vivo SAMase activity were assayed for expression of the metA, B, C, E, F, H, J, K and R genes in cells grown in methionine-rich complete media as well as in defined media with and without l-methionine. In vivo SAMase activity dramatically induced expression between 11- and nearly 1000-fold depending on the gene assayed for all but metJ and metH, and these genes were induced over twofold. metJ : : Tn5 (aporepressor defective) and metK : : Tn5 (SAM synthetase impaired; produces <5 % of wild-type SAM) strains containing in vivo SAMase activity produced even higher met gene activity than that seen in comparably prepared cells with wild-type genes for all but metJ in a MetJ-deficient background. The SAMase-mediated hyperinduction of metH in wild-type cells and of the met genes assayed in metJ : : Tn5 and metK : : Tn5 cells provokes questions about how other elements such as the MetR activator protein or factors beyond the met regulon itself might be involved in the regulation of genes responsible for methionine biosynthesis.
LaMonte
Bernadette L
BL
Department of Biology, Ursinus College, PO Box 1000, Collegeville, PA 19426, USA.
Hughes
Jeffrey A
JA
Biology Department, Hanover College, PO Box 890, Hanover, IN 47243, USA.
D019677:Artificial Gene Fusion; D017903:Bacteriophage T3 / Q000235:genetics; D004251:DNA Transposable Elements / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000235:genetics; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter / Q000235:genetics / Q000502:physiology; D006867:Hydrolases / Q000096:biosynthesis / Q000235:genetics; D006868:Hydrolysis; D008715:Methionine / Q000096:biosynthesis*; D016254:Mutagenesis, Insertional; D010957:Plasmids / Q000235:genetics; D018085:Regulon / Q000502:physiology*; D012436:S-Adenosylmethionine / Q000378:metabolism*; D014169:Transformation, Bacterial; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D004251:DNA Transposable Elements; D029968:Escherichia coli Proteins; D014764:Viral Proteins; D012436:S-Adenosylmethionine; D008715:Methionine; D006867:Hydrolases; D001616:beta-Galactosidase; C019614:adenosylmethionine hydrolase