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Volume 149, Issue 8

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

Abstract

The gene, encoding the enzyme polynucleotide phosphorylase (PNPase), was overexpressed in the actinomycin producer . Integration of pIJ8600, bearing the thiostrepton-inducible promoter, and its derivatives containing into the chromosome dramatically increased the growth rate of the resulting strains as compared with the parent strain. Thiostrepton induction of a strain containing pJSE340, bearing 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 ORF and some 3′-flanking sequence, led to lower levels of PNPase activity and a different pattern of expression compared with pJSE340. Induction of 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 as compared with controls but it was not possible to attribute this decrease specifically to the increase in PNPase levels. Overexpression of had no effect on ppGpp levels in the relevant strains. It was observed that the 3′-tails associated with RNAs from are heteropolymeric. The authors argue that those tails are synthesized by PNPase rather than by a poly(A) polymerase similar to that found in and that PNPase may be the sole RNA 3′-polynucleotide polymerase in streptomycetes.

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Keyword(s): acm, actinomycin D and PNPase, polynucleotide phosphorylase
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2003-08-01
2024-04-19
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Overexpression of the polynucleotide phosphorylase gene (pnp) of Streptomyces antibioticus affects mRNA stability and poly(A) tail length but not ppGpp levels
Microbiology 149, 2173 (2003); https://doi.org/10.1099/mic.0.26334-0
/content/journal/micro/10.1099/mic.0.26334-0
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