Probing the substrate specificity of the bacterial Pnkp/Hen1 RNA repair system using synthetic RNAs

Can Zhang, Chio Mui Chan, Pei Wang, Raven H. Huang

Research output: Contribution to journalArticle

Abstract

Ribotoxins cleave essential RNAs involved in protein synthesis as a strategy for cell killing. RNA repair systems exist in nature to counteract the lethal actions of ribotoxins, as first demonstrated by the RNA repair system from bacteriophage T4 25 yr ago. Recently, we found that two bacterial proteins, named Pnkp and Hen1, form a stable complex and are able to repair ribotoxin-cleaved tRNAs in vitro. However, unlike the well-studied T4 RNA repair system, the natural RNA substrates of the bacterial Pnkp/Hen1 RNA repair system are unknown. Here we present comprehensive RNA repair assays with the recombinant Pnkp/Hen1 proteins from Anabaena variabilis using a total of 33 different RNAs as substrates that might mimic various damaged forms of RNAs present in living cells. We found that unlike the RNA repair system from bacteriophage T4, the bacterial Pnkp/Hen1 RNA repair system exhibits broad substrate specificity. Based on the experimental data presented here, a model of preferred RNA substrates of the Pnkp/Hen1 repair system is proposed.

Original languageEnglish (US)
Pages (from-to)335-344
Number of pages10
JournalRNA
Volume18
Issue number2
DOIs
StatePublished - Feb 1 2012

Keywords

  • Pnkp/Hen1
  • RNA damage
  • RNA repair
  • Ribotoxin
  • Substrate specificity

ASJC Scopus subject areas

  • Molecular Biology

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