In vivo 3'-to-5' exoribonuclease targetomes of Streptococcus pyogenes

Proc Natl Acad Sci U S A. 2018 Nov 13;115(46):11814-11819. doi: 10.1073/pnas.1809663115. Epub 2018 Oct 31.

Abstract

mRNA decay plays an essential role in the control of gene expression in bacteria. Exoribonucleases (exoRNases), which trim transcripts starting from the 5' or 3' end, are particularly important to fully degrade unwanted transcripts and renew the pool of nucleotides available in the cell. While recent techniques have allowed genome-wide identification of ribonuclease (RNase) targets in bacteria in vivo, none of the 3'-to-5' exoRNase targetomes (i.e., global processing sites) have been studied so far. Here, we report the targetomes of YhaM, polynucleotide phosphorylase (PNPase), and RNase R of the human pathogen Streptococcus pyogenes We determined that YhaM is an unspecific enzyme that trims a few nucleotides and targets the majority of transcript ends, generated either by transcription termination or by endonucleolytic activity. The molecular determinants for YhaM-limited processivity are yet to be deciphered. We showed that PNPase clears the cell from mRNA decay fragments produced by endoribonucleases (endoRNases) and is the major 3'-to-5' exoRNase for RNA turnover in S. pyogenes In particular, PNPase is responsible for the degradation of regulatory elements from 5' untranslated regions. However, we observed little RNase R activity in standard culture conditions. Overall, our study sheds light on the very distinct features of S. pyogenes 3'-to-5' exoRNases.

Keywords: 3′-end sequencing; 3′-to-5′ exoRNase; 5′-end sequencing; RNA degradation; Streptococcus pyogenes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Exoribonucleases / metabolism*
  • Exoribonucleases / physiology
  • Gene Expression Regulation, Bacterial / genetics
  • Polyribonucleotide Nucleotidyltransferase / metabolism
  • RNA Stability / genetics
  • RNA Stability / physiology*
  • RNA, Bacterial / genetics
  • RNA, Messenger / metabolism
  • Streptococcus pyogenes / genetics*

Substances

  • RNA, Bacterial
  • RNA, Messenger
  • Polyribonucleotide Nucleotidyltransferase
  • Exoribonucleases
  • ribonuclease R