PMID- 22101242
OWN - NLM
STAT- MEDLINE
DCOM- 20120206
LR  - 20211021
IS  - 1469-9001 (Electronic)
IS  - 1355-8382 (Print)
IS  - 1355-8382 (Linking)
VI  - 18
IP  - 1
DP  - 2012 Jan
TI  - Determinants of the cytotoxicity of PrrC anticodon nuclease and its amelioration 
      by tRNA repair.
PG  - 145-54
LID - 10.1261/rna.030171.111 [doi]
AB  - Breakage of tRNA(Lys(UUU)) by the Escherichia coli anticodon nuclease PrrC 
      (EcoPrrC) underlies a host antiviral response to phage T4 infection that is 
      ultimately thwarted by a virus-encoded RNA repair system. PrrC homologs are 
      prevalent in other bacteria, but their activities and substrates are not defined. 
      We find that induced expression of EcoPrrC is toxic in Saccharomyces cerevisiae 
      and E. coli, whereas the Neisseria meningitidis PrrC (NmePrrC) is not. PrrCs 
      consist of an N-terminal NTPase module and a C-terminal nuclease module. Domain 
      swaps identified the EcoPrrC nuclease domain as decisive for toxicity when linked 
      to either the Eco or Nme NTPase. Indeed, a single arginine-to-tryptophan change 
      in the NmePrrC nuclease domain (R316W) educed a gain-of-function and rendered 
      NmePrrC toxic to yeast, with genetic evidence for tRNA(Lys(UUU)) being the 
      relevant target. The reciprocal Trp-to-Arg change in EcoPrrC (W335R) abolished 
      its toxicity. Further mutagenesis of the EcoPrrC nuclease domain highlighted an 
      ensemble of 15 essential residues and distinguished between hypomorphic alleles 
      and potential nuclease-nulls. We report that the RNA repair phase of the 
      bacterial virus-host dynamic is also portable to yeast, where coexpression of the 
      T4 enzymes Pnkp and Rnl1 ameliorated the toxicity of NmePrrC-R316W. Plant tRNA 
      ligase AtRNL also countered NmePrrC-R316W toxicity, in a manner that depended on 
      AtRNL's 5'-kinase and ligase functions.
FAU - Meineke, Birthe
AU  - Meineke B
AD  - Molecular Biology Program, Sloan-Kettering Institute, New York, New York 10065, 
      USA.
FAU - Shuman, Stewart
AU  - Shuman S
LA  - eng
GR  - R01 GM042498/GM/NIGMS NIH HHS/United States
GR  - GM42498/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20111118
PL  - United States
TA  - RNA
JT  - RNA (New York, N.Y.)
JID - 9509184
RN  - 0 (Escherichia coli Proteins)
RN  - 8DUH1N11BX (Tryptophan)
RN  - 9014-25-9 (RNA, Transfer)
RN  - 94ZLA3W45F (Arginine)
RN  - EC 3.1.- (PrrC protein, E coli)
RN  - EC 3.1.- (Ribonucleases)
RN  - EC 3.1.- (anticodon nuclease)
RN  - WHI7HQ7H85 (Uridine)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Arginine/chemistry/genetics
MH  - Escherichia coli Proteins/*chemistry/genetics
MH  - Gene Dosage
MH  - Molecular Sequence Data
MH  - Neisseria meningitidis/*enzymology
MH  - Protein Structure, Tertiary
MH  - RNA, Transfer/*chemistry/genetics
MH  - Ribonucleases/*chemistry/genetics
MH  - Saccharomyces cerevisiae/enzymology/genetics
MH  - Tryptophan/chemistry/genetics
MH  - Uridine/chemistry
PMC - PMC3261736
EDAT- 2011/11/22 06:00
MHDA- 2012/02/07 06:00
PMCR- 2013/01/01
CRDT- 2011/11/22 06:00
PHST- 2011/11/22 06:00 [entrez]
PHST- 2011/11/22 06:00 [pubmed]
PHST- 2012/02/07 06:00 [medline]
PHST- 2013/01/01 00:00 [pmc-release]
AID - rna.030171.111 [pii]
AID - RA [pii]
AID - 10.1261/rna.030171.111 [doi]
PST - ppublish
SO  - RNA. 2012 Jan;18(1):145-54. doi: 10.1261/rna.030171.111. Epub 2011 Nov 18.