ADP-ribosylation of proteins in Bacillus subtilis and its possible importance in sporulation.
Endogenous ADP-ribosylation was detected in Bacillus subtilis, as determined in vitro with crude cellular extracts. The ADP-ribosylated protein profile changed during growth in sporulation medium, displaying a temporary appearance of two ADP-ribosylated proteins (36 and 58 kDa) shortly after the end of exponential growth. Mutants resistant to 3-methoxybenzamide, a known inhibitor of ADP-ribosyltransferase, were obtained, and a significant proportion (15%) were found to be defective in both sporulation and antibiotic production. These mutants failed to ADP-ribosylate the 36- and 58-kDa proteins. The parent strain also lost the ability to ADP-ribosylate these proteins when grown in the presence of 3-methoxybenzamide at a concentration at which sporulation but not cell growth was severely inhibited. Results from genetic transformations showed that the mutation conferring resistance to 3-methoxybenzamide, named brgA, was cotransformed with the altered phenotypes, i.e., defects in ADP-ribosylation and sporulation. spoOA and spoOF mutants displayed an ADP-ribosylation profile similar to that of the parent strain, but a spoOH mutant failed to ADP-ribosylate any proteins, including the 36- and 58-kDa proteins. The significance of protein ADP-ribosylation in sporulation was further indicated by the observation that ADP-ribosylation of the 36-kDa protein could be induced by treatment with decoyinine, an inhibitor of GMP-synthetase, and by amino acid limitation, both of which resulted in an immediate decrease in GTP pool size eventually leading to massive sporulation. We propose that a new sporulation gene, which presumably controls sporulation via ADP-ribosylation of certain functional proteins, exists.
Huh
J W
JW
National Food Research Institute, Ibaraki, Japan.
Shima
J
J
Ochi
K
K
D000246:Adenosine Diphosphate Ribose / Q000378:metabolism*; D001412:Bacillus subtilis / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D001549:Benzamides / Q000494:pharmacology; D004352:Drug Resistance, Microbial; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D008970:Molecular Weight; D016296:Mutagenesis; D009243:NAD / Q000378:metabolism*; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D013171:Spores, Bacterial
D001426:Bacterial Proteins; D001549:Benzamides; D004791:Enzyme Inhibitors; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D009243:NAD; D000246:Adenosine Diphosphate Ribose; D011065:Poly(ADP-ribose) Polymerases; C025159:3-methoxybenzamide