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Department of Biology, 1510 Clifton Rd, Emory University, Atlanta, GA 30322, USA
Correspondence
G. H. Jones
gjones{at}biology.emory.edu
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.
This article has been cited by other articles:
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  S. A. Chang, M. Cozad, G. A. Mackie, and G. H. Jones Kinetics of Polynucleotide Phosphorylase: Comparison of Enzymes from Streptomyces and Escherichia coli and Effects of Nucleoside Diphosphates J. Bacteriol., January 1, 2008; 190(1): 98 - 106. [Abstract] [Full Text] [PDF]
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 P. Bralley, B. Gust, S. Chang, K. F. Chater, and G. H. Jones RNA 3'-tail synthesis in Streptomyces: in vitro and in vivo activities of RNase PH, the SCO3896 gene product and polynucleotide phosphorylase. Microbiology, March 1, 2006; 152(Pt 3): 627 - 636. [Abstract] [Full Text] [PDF]
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P. Bralley, S. A. Chang, and G. H. Jones A Phylogeny of Bacterial RNA Nucleotidyltransferases: Bacillus halodurans Contains Two tRNA Nucleotidyltransferases J. Bacteriol., September 1, 2005; 187(17): 5927 - 5936. [Abstract] [Full Text] [PDF]
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J. Campos-Guillen, P. Bralley, G. H. Jones, D. H. Bechhofer, and G. Olmedo-Alvarez Addition of Poly(A) and Heteropolymeric 3' Ends in Bacillus subtilis Wild-Type and Polynucleotide Phosphorylase-Deficient Strains J. Bacteriol., July 15, 2005; 187(14): 4698 - 4706. [Abstract] [Full Text] [PDF]
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 T. Nagaike, T. Suzuki, T. Katoh, and T. Ueda Human Mitochondrial mRNAs Are Stabilized with Polyadenylation Regulated by Mitochondria-specific Poly(A) Polymerase and Polynucleotide Phosphorylase J. Biol. Chem., May 20, 2005; 280(20): 19721 - 19727. [Abstract] [Full Text] [PDF]
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P. Bralley and G. H. Jones Organization and Expression of the Polynucleotide Phosphorylase Gene (pnp) of Streptomyces: Processing of pnp Transcripts in Streptomyces antibioticus J. Bacteriol., May 15, 2004; 186(10): 3160 - 3172. [Abstract] [Full Text] [PDF]
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