Protein-only RNase P function in Escherichia coli: viability, processing defects and differences between PRORP isoenzymes

Nucleic Acids Research
Markus GössringerR K Hartmann

Abstract

The RNase P family comprises structurally diverse endoribonucleases ranging from complex ribonucleoproteins to single polypeptides. We show that the organellar (AtPRORP1) and the two nuclear (AtPRORP2,3) single-polypeptide RNase P isoenzymes from Arabidopsis thaliana confer viability to Escherichia coli cells with a lethal knockdown of its endogenous RNA-based RNase P. RNA-Seq revealed that AtPRORP1, compared with bacterial RNase P or AtPRORP3, cleaves several precursor tRNAs (pre-tRNAs) aberrantly in E. coli. Aberrant cleavage by AtPRORP1 was mainly observed for pre-tRNAs that can form short acceptor-stem extensions involving G:C base pairs, including tRNAAsp(GUC), tRNASer(CGA) and tRNAHis. However, both AtPRORP1 and 3 were defective in processing of E. coli pre-tRNASec carrying an acceptor stem expanded by three G:C base pairs. Instead, pre-tRNASec was degraded, suggesting that tRNASec is dispensable for E. coli under laboratory conditions. AtPRORP1, 2 and 3 are also essentially unable to process the primary transcript of 4.5S RNA, a hairpin-like non-tRNA substrate processed by E. coli RNase P, indicating that PRORP enzymes have a narrower, more tRNA-centric substrate spectrum than bacterial RNA-based RNase P enzymes. The cel...Continue Reading

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Citations

Oct 27, 2017·Proceedings of the National Academy of Sciences of the United States of America·Astrid I NickelRoland K Hartmann
Oct 4, 2017·RNA·Venkat GopalanAndrey S Krasilnikov
Jun 28, 2018·Amino Acids·Vitor Hugo Balasco SerrãoOtavio Henrique Thiemann
Aug 30, 2019·Critical Reviews in Biochemistry and Molecular Biology·David H Bechhofer, Murray P Deutscher
Mar 8, 2019·Wiley Interdisciplinary Reviews. RNA·Vincent BoivinSherif Abou-Elela
Sep 7, 2021·Molecular Microbiology·Bijoy K Mohanty, Sidney R Kushner

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Datasets Mentioned

BETA
CP012868.1
and

Methods Mentioned

BETA
RNA-Seq
PCR
aminoacylation

Software Mentioned

FASTX Toolkit
custom
segemehl

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