A rifampicin resistance mutation in the rpoB gene confers ppGpp-independent antibiotic production in Streptomyces coelicolor A3(2).
In Streptomyces coelicolor A3(2), deletion of relA or a specific mutation in rplK ( relC) results in an inability to synthesize ppGpp (guanosine 5'-diphosphate 3'-diphosphate) and impairs production of actinorhodin. We have found that certain rifampicin-resistant ( rif) mutants isolated from either relA or relC strains regain the ability to produce actinorhodin at the same level as the wild-type strain, although their capacity to synthesize ppGpp is unchanged. These rif mutants were found to have a missense mutation in the rpoB gene that encodes the RNA polymerase beta-subunit. This rpoB mutation was shown to be responsible for the observed changes in phenotype, as demonstrated by gene replacement experiments. Gene expression analysis revealed that the restoration of actinorhodin production in both relA and relC strains is accompanied by increased expression of the pathway-specific regulator gene actII-ORF4, which is normally decreased in the rel mutants. In addition to the restoration of antibiotic production, the rif mutants also exhibited a lower rate of RNA synthesis compared to the parental strain when grown in a rich medium, suggesting that these mutant RNA polymerases behave like "stringent" RNA polymerases. These results indicate that rif mutations can alter gene expression patterns independently of ppGpp. We propose that RNA polymerases carrying particular rif mutations in the beta-subunit can functionally mimic the modification induced by binding of ppGpp.
Xu
J
J
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Tozawa
Y
Y
Lai
C
C
Hayashi
H
H
Ochi
K
K
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000494:pharmacology; D000880:Anthraquinones / Q000378:metabolism; D001426:Bacterial Proteins; D001665:Binding Sites; D012321:DNA-Directed RNA Polymerases / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D004768:Enterotoxins / Q000378:metabolism; D006150:Guanine Nucleotides / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D009154:Mutation; D012293:Rifampin / Q000494:pharmacology*; D013302:Streptomyces / Q000235:genetics*; D000095622:Benzoisochromanequinones
D000596:Amino Acids; D000880:Anthraquinones; D001426:Bacterial Proteins; D012321:DNA-Directed RNA Polymerases; D004768:Enterotoxins; D006150:Guanine Nucleotides; D012293:Rifampin; C017721:casamino acids; C082174:cytolytic enterotoxin protein, Aeromonas; C018957:guanosine 5'-diphosphate-3'-monophosphate; C105078:RNA polymerase beta subunit; C013390:actinorhodin; D000095622:Benzoisochromanequinones