Second-site suppression of RNase E essentiality by mutation of the deaD RNA helicase in Escherichia coli.

Journal of Bacteriology
Masaru TamuraS N Cohen

Abstract

Escherichia coli cells normally require RNase E activity to propagate and form colonies. Using random Tn10 insertion mutagenesis, we screened for second-site suppressor mutations that restore colony-forming ability (CFA) to E. coli cells lacking RNase E function and found mutations in three separate chromosomal loci that had this phenotype. Restoration of CFA by mutations in two of the genes identified was observed only in nutrient-poor medium, whereas the effects of mutation of the ATP-dependent RNA helicase DeaD were medium independent. Suppression of the rne mutant phenotype by inactivation of deaD was partial, as rne deaD doubly mutant bacteria had a greatly prolonged generation time and grew as filamentous chains in liquid medium. Moreover, we found that CFA restoration by deaD inactivation requires normal expression of the endogenous rng gene in doubly mutant rne deaD cells. Second-site suppression by deaD mutation was attributable specifically to ablation of the helicase activity of DeaD and was reversed by adventitious expression of RhlE or RNase R, both of which can unwind double-stranded RNA. Our results suggest a previously unsuspected role for RNA secondary structure as a determinant of RNase E essentiality.

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Citations

Sep 21, 2013·The Journal of Biological Chemistry·Zbigniew PietrasBen F Luisi
Jan 1, 2013·Journal of Bacteriology·Masaru TamuraStanley N Cohen
Sep 26, 2013·Cellular and Molecular Life Sciences : CMLS·Soumaya LaalamiHarald Putzer
Feb 18, 2016·Archives of Microbiology·Masaru TamuraAtsushi Kato
Jan 26, 2016·Current Opinion in Microbiology·Vanessa Khemici, Patrick Linder
Apr 25, 2015·FEMS Microbiology Reviews·Peter RedderPatrick Linder
Feb 14, 2013·Molecular BioSystems·Li ZhouChristine Vogel
Dec 6, 2019·Nucleic Acids Research·Nida Ali, Jayaraman Gowrishankar

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