Absence of RNASE III alters the pathway by which RNAI, the antisense inhibitor of ColE1 replication, decays

Uta Binnie, Kenny Wong, Sean McAteer, Millicent/Mrs Donachie Masters

Research output: Contribution to journalArticlepeer-review

Abstract

RNAI is a short RNA, 108 nt in length, which regulates the replication of the plasmid ColE1. RNAI turns over rapidly, enabling plasmid replication rate to respond quickly to changes in plasmid copy number. Because RNAI is produced in abundance, is easily extracted and turns over quickly, it has been used as a model for mRNA in studying RNA decay pathways. The enzymes polynucleotide phosphorylase, poly(A) polymerase and RNase E have been demonstrated to have roles in both messenger and RNAI decay; it is reported here that these enzymes can work independently of one another to facilitate RNAI decay. The roles in RNAI decay of two further enzymes which facilitate mRNA decay, the exonuclease RNase II and the endonuclease RNase III, are also examined. RNase II does not appear to accelerate RNAI decay but it is found that, in the absence of RNase III, polyadenylated RNAI, unprocessed by RNase E, accumulates. It is also shown that RNase III can cut RNAI near nt 82 or 98 in vitro. An RNAI fragment corresponding to the longer of these can be found in extracts of an mc+ pcnB strain (which produces RNase III) but not of an rnc pcnB strain, suggesting that RNAI may be a substrate for RNase III in vivo. A possible pathway for the early steps in RNAI decay which incorporates this information is suggested.
Original languageEnglish
Article number11
Pages (from-to)3089-100
Number of pages12
JournalMicrobiology
Volume145
Publication statusPublished - Nov 1999

Keywords / Materials (for Non-textual outputs)

  • Bacterial Proteins
  • Blotting, Northern
  • DNA Replication/drug effects
  • Endoribonucleases/genetics
  • Endoribonucleases/physiology
  • Escherichia coli/enzymology
  • Escherichia coli Proteins
  • Exoribonucleases/genetics
  • Exoribonucleases/physiology
  • Mutation
  • Plasmids/physiology
  • Polymerase Chain Reaction
  • Polynucleotide Adenylyltransferase/genetics
  • Polynucleotide Adenylyltransferase/physiology
  • Protein Processing, Post-Translational
  • RNA, Antisense/metabolism
  • RNA, Antisense/pharmacology
  • RNA, Bacterial/metabolism
  • RNA, Messenger/metabolism
  • RNA, Small Interfering

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