Expression of the cloned coliphage T3 S-adenosylmethionine hydrolase gene inhibits DNA methylation and polyamine biosynthesis in Escherichia coli.
We have developed a new research tool for the study of S-adenosylmethionine (AdoMet) metabolism by cloning the coliphage T3 AdoMet hydrolase (AdoMetase; EC 3.3.1.2) gene into the M13mp8 expression vector. The recombinant bacteriophage clones expressed an AdoMetase activity in Escherichia coli like that found in T3-infected cells. High levels of AdoMetase expression impaired AdoMet-mediated activities such as dam and dcm methylase-directed DNA modifications and the synthesis of spermidine from putrescine. Expression vectors containing the cloned AdoMetase gene thus provide an alternate approach to the use of chemical inhibitors or mutants defective in AdoMet biosynthesis to probe the effect of AdoMet limitation.
Hughes
J A
JA
Brown
L R
LR
Ferro
A J
AJ
D003001:Cloning, Molecular; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000378:metabolism*; D005814:Genes, Viral; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism*; D008745:Methylation; D012436:S-Adenosylmethionine / Q000378:metabolism*; D013095:Spermidine / Q000096:biosynthesis*; D013604:T-Phages / Q000201:enzymology / Q000235:genetics
D004269:DNA, Bacterial; D012436:S-Adenosylmethionine; D004248:DNA (Cytosine-5-)-Methyltransferases; D006867:Hydrolases; C019614:adenosylmethionine hydrolase; D013095:Spermidine