SCO5745, a Bifunctional RNase J Ortholog, Affects Antibiotic Production in Streptomyces coelicolor

The bacterial RNases J are considered bifunctional RNases possessing both endo- and exonucleolytic activities. We have isolated an RNase J ortholog from Streptomyces coelicolor encoded by the gene sco5745. We overexpressed a decahistidine-tagged version of SCO5745 and purified the overexpressed prot...

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Bibliographic Details
Published in:Journal of Bacteriology 2014-03, Vol.196 (6), p.1197-1205
Main Authors: Bralley, Patricia, Aseem, Madiha, Jones, George H
Format: Article
Language:English
Subjects:
affinity chromatography
alleles
Anthraquinones - metabolism
Anti-Bacterial Agents - biosynthesis
Antibiotics
Bacillus subtilis
Bacteriology
Chromatography
Chromatography, Affinity
Gene Expression
Gene Expression Regulation, Bacterial
Gene Knockout Techniques
Gram-positive bacteria
Hydrolysis
Prodigiosin - analogs & derivatives
Prodigiosin - biosynthesis
Proteins
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
ribonucleases
Ribonucleases - genetics
Ribonucleases - isolation & purification
Ribonucleases - metabolism
Ribonucleic acid
RNA
RNA, Messenger - metabolism
Streptomyces coelicolor
Streptomyces coelicolor - genetics
Streptomyces coelicolor - metabolism
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Summary:The bacterial RNases J are considered bifunctional RNases possessing both endo- and exonucleolytic activities. We have isolated an RNase J ortholog from Streptomyces coelicolor encoded by the gene sco5745. We overexpressed a decahistidine-tagged version of SCO5745 and purified the overexpressed protein by immobilized metal ion affinity chromatography. We demonstrated the presence of both 5′-to-3′ exonucleolytic and endonucleolytic activities on the Bacillus subtilis thrS transcript. Exonucleoytic activity predominated with 5′ monophosphorylated thrS, while endonucleolytic activity predominated with 5′ triphosphorylated thrS. While sco5745 is the only RNase J allele in S. coelicolor, the gene is not essential. Its disruption resulted in delayed production of the antibiotic actinorhodin, overproduction of undecylprodigiosin, and diminished production of the calcium-dependent antibiotic, in comparison with the parental strain.
ISSN:0021-9193
1098-5530
1067-8832
DOI:10.1128/JB.01422-13