Genetic analysis of absB, a Streptomyces coelicolor locus involved in global antibiotic regulation

Adamidis, Trifon; Champness, Wendy

doi:10.1128/jb.174.14.4622-4628.1992

Cited by 53 publications

(64 citation statements)

References 34 publications

(45 reference statements)

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“…An effect similar to that observed with the actII-ORF4 gene has been described previously in absA and absB mutants (91,92). These data also support the suggestion that the ActII-ORF4 protein is by itself sufficient to activate transcription of the biosynthetic genes, and if any other additional factor is required, either it does not constitute a limitation or its action might be overtaken by the overproduced ActII-ORF4 protein (93).…”

Section: (P)ppgpp Synthetase From S Coelicolorsupporting

confidence: 74%

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

Martı́nez-Costa

Arias

Romero

et al. 1996

Journal of Biological Chemistry

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A 0.972-kilobase pair DNA fragment from Streptomyces lividans that induces the production of the bluepigmented antibiotic actinorhodine in S. lividans when cloned on a multicopy plasmid has led to the isolation of a 4-kilobase pair DNA fragment from Streptomyces coelicolor containing homologous sequence. Computer-assisted analysis of the DNA sequence revealed three putative open reading frames (ORFs), ORF1, ORF2, and ORF3. ORF2 extends beyond the sequenced DNA fragment, and its deduced product shares no similarities with any other known proteins in the data bases. ORF3 is also truncated, and its 41-amino acid C-terminal product is identical to the S. coelicolor adenine phosphoribosyltransferase. The 847-amino acid ORF1 protein, with a predicted molecular mass of 94.2 kDa, strongly resembled the relA and spoT gene products from Escherichia coli and the homologs from Vibrio sp. strain S14, Haemophilus influenzae, Streptococcus equisimilis H46A, and Mycoplasma genitalium. Unlike these proteins, the ORF1 amino acid sequence analysis revealed the presence of a putative ATP/GTP-binding domain. A mutant was generated by deleting most of the ORF1 gene that showed an actinorhodine-nonproducing phenotype, while undecylprodigiosin and the calcium-dependent antibiotic were unaffected. The mutant strain grew at a much lower rate than the wild-type strain, and spore formation was delayed. When the gene was propagated on a low copy number vector, not only was actinorhodine production restored, but actinorhodine and undecylprodigiosin production was enhanced in both the mutant and wild-type strains and morphological differentiation returned to wild-type characteristics. (p)ppGpp synthetase activity was not detected in purified ribosomes from the ORF1-deleted mutant, while it was restored by complementation of this strain.

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Section: (P)ppgpp Synthetase From S Coelicolorsupporting

confidence: 74%

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

Martı́nez-Costa

Arias

Romero

et al. 1996

Journal of Biological Chemistry

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“…The absA and absB mutations result in a global block of antibiotic production in S. coelicolor (1,2). absA and absB were mapped at 10 and 4 o'clock, respectively, on the chromosomal linkage map (Fig.…”

Section: Resultsmentioning

confidence: 99%

A putative two-component regulatory system involved in secondary metabolism in Streptomyces spp

et al. 1992

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A DNA fragment stimulating actinorhodin, undecylprodigiosin, and A-factor production in Streptomyces lividans 66 was cloned from Streptomyces coelicolor A3(2). Nucleotide sequencing revealed the presence of an open reading frame of 225 codons, named afsQl, that showed great similarity in amino acid sequence to the response regulators of typical prokaryotic two-component regulatory systems responsible for adaptive responses. The termination codon, TGA, of afsQl overlapped the initiation codon, GTG, of a second open reading frame, afsQ2, of 535 codons. The afsQ2 gene product showed homology with the sensory histidine protein kinases of two-component systems. In agreement with the assumption that the AfsQl and AfsQ2 proteins comprise an aspartate-histidine phosphotransfer system, an amino acid replacement from Asp to Glu at residue 52 of AfsQl, generated by site-directed mutagenesis, resulted in loss of the protein's ability to stimulate antibiotic production in S. lividans. Primer extension experiments indicated that transcription of the afsQl and afsQ2 genes initiates at the translational start codon (GTG) of the afsQl gene. The afsQ1 and afsQ2 genes were physically mapped at a chromosomal position near the actinorhodin biosynthetic gene cluster (act) by hybridization to Southern blots of restriction fragments separated by pulsed-field gel electrophoresis.

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“…A gene encoding a putative RNase III homologue was identified as the absB locus in S. coelicolor (Adamis & Champness, 1992;Price et al, 1999). Interestingly, mutations in absB abolish production of all four antibiotics normally formed by S. coelicolor (Adamis & Champness, 1992), suggesting a possible relationship between RNA degradation and the regulation of antibiotic production in Abbreviations: acm, actinomycin D; PNPase, polynucleotide phosphorylase.…”

Section: Introductionmentioning

confidence: 99%

Overexpression of the polynucleotide phosphorylase gene (pnp) of Streptomyces antibioticus affects mRNA stability and poly(A) tail length but not ppGpp levels

Bralley

Jones

2003

Microbiology

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The pnp gene, encoding the enzyme polynucleotide phosphorylase (PNPase), was overexpressed in the actinomycin producer Streptomyces antibioticus. Integration of pIJ8600, bearing the thiostrepton-inducible tipA promoter, and its derivatives containing pnp into the S. antibioticus chromosome dramatically increased the growth rate of the resulting strains as compared with the parent strain. Thiostrepton induction of a strain containing pJSE340, bearing pnp with a 5′-flanking region containing an endogenous promoter, led to a 2·5–3 fold increase in PNPase activity levels, compared with controls. Induction of a strain containing pJSE343, with only the pnp ORF and some 3′-flanking sequence, led to lower levels of PNPase activity and a different pattern of pnp expression compared with pJSE340. Induction of pnp from pJSE340 resulted in a decrease in the chemical half-life of bulk mRNA and a decrease in poly(A) tail length as compared to RNAs from controls. Actinomycin production decreased in strains overexpressing pnp as compared with controls but it was not possible to attribute this decrease specifically to the increase in PNPase levels. Overexpression of pnp had no effect on ppGpp levels in the relevant strains. It was observed that the 3′-tails associated with RNAs from S. antibioticus are heteropolymeric. The authors argue that those tails are synthesized by PNPase rather than by a poly(A) polymerase similar to that found in Escherichia coli and that PNPase may be the sole RNA 3′-polynucleotide polymerase in streptomycetes.

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Genetic analysis of absB, a Streptomyces coelicolor locus involved in global antibiotic regulation

Cited by 53 publications

References 34 publications

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

A relA/spoT Homologous Gene from Streptomyces coelicolor A3(2) Controls Antibiotic Biosynthetic Genes

A putative two-component regulatory system involved in secondary metabolism in Streptomyces spp

Overexpression of the polynucleotide phosphorylase gene (pnp) of Streptomyces antibioticus affects mRNA stability and poly(A) tail length but not ppGpp levels

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