SCO5745, a Bifunctional RNase J Ortholog, affects antibiotic production in streptomyces coelicolor

• Type of Material: Article
• Authors: Patricia Bralley, Emory University; Madiha Aseem, Emory University; George Jones, Emory University
• Language: English
• Date: 2014-03-01
• Publisher: American Society for Microbiology
• Publication Version: Final Published Version
• Copyright Statement: © 2014, American Society for Microbiology.
• Final Published Version (URL): http://dx.doi.org/10.1128/JB.01422-13
• Title of Journal or Parent Work: Journal of Bacteriology
• ISSN: 0021-9193
• Volume: 196
• Issue: 6
• Start Page: 1197
• End Page: 1205
• Grant/Funding Information: This research was supported by grant MCB 0817177 from the National Science Foundation to G.H.J.
• Keywords: Gene Expression; Recombinant Proteins; RNA, Messenger; Gene Expression Regulation, Bacterial; Anti-Bacterial Agents; Prodigiosin; Gene Knockout Techniques; Ribonucleases; Chromatography, Affinity; Hydrolysis; Anthraquinones; Streptomyces coelicolor
• Research Categories: Biology, General; Biology, Physiology

Abstract

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.