PMID- 20417643
OWN - NLM
STAT- MEDLINE
DCOM- 20100908
LR  - 20231127
IS  - 1089-8638 (Electronic)
IS  - 0022-2836 (Print)
IS  - 0022-2836 (Linking)
VI  - 399
IP  - 5
DP  - 2010 Jun 25
TI  - Mechanism of U-insertion RNA editing in trypanosome mitochondria: 
      characterization of RET2 functional domains by mutational analysis.
PG  - 696-706
LID - 10.1016/j.jmb.2010.04.035 [doi]
AB  - 3'-Terminal uridylyl transferases (TUTases) selectively bind uridine 
      5'-triphosphate (UTP) and catalyze the addition of uridine 5'-monophosphate to 
      the 3'-hydroxyl of RNA substrates in a template-independent manner. RNA editing 
      TUTase 1 and RNA editing TUTase 2 (RET2) play central roles in uridine 
      insertion/deletion RNA editing, which is an essential part of mitochondrial RNA 
      processing in trypanosomes. Although the conserved N-terminal (catalytic) domain 
      and C-terminal (nucleotide base recognition) domain are readily distinguished in 
      all known TUTases, nucleotide specificity, RNA substrate preference, 
      processivity, quaternary structures, and auxiliary domains vary significantly 
      among enzymes of divergent biological functions. RET2 acts as a subunit of the 
      RNA editing core complex to carry out guide-RNA-dependent U-insertion into 
      mitochondrial mRNA. By correlating mutational effects on RET2 activity as 
      recombinant protein and as RNA editing core complex subunit with RNAi-based 
      knock-in phenotypes, we have assessed the UTP and RNA binding sites in RET2. Here 
      we demonstrate functional conservation of key UTP-binding and 
      metal-ion-coordinating residues and identify amino acids involved in RNA 
      substrate recognition. Invariant arginine residues 144 and 435 positioned in the 
      vicinity of the UTP binding site are critical for RET2 activity on 
      single-stranded and double-stranded RNAs, as well as function in vivo. 
      Recognition of a double-stranded RNA, which resembles a guide RNA/mRNA duplex, is 
      further facilitated by multipoint contacts across the RET2-specific middle 
      domain.
CI  - Copyright (c) 2010 Elsevier Ltd. All rights reserved.
FAU - Ringpis, Gene-Errol
AU  - Ringpis GE
AD  - Department of Microbiology and Molecular Genetics, School of Medicine, University 
      of California Irvine, B240 Medical Sciences I, Irvine, CA 92697, USA.
FAU - Stagno, Jason
AU  - Stagno J
FAU - Aphasizhev, Ruslan
AU  - Aphasizhev R
LA  - eng
GR  - R01 AI064653/AI/NIAID NIH HHS/United States
GR  - R01 AI064653-05/AI/NIAID NIH HHS/United States
GR  - AI064653/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20100424
PL  - Netherlands
TA  - J Mol Biol
JT  - Journal of molecular biology
JID - 2985088R
RN  - 0 (Protozoan Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Mitochondrial)
RN  - 0 (RNA, Protozoan)
RN  - 0 (mitochondrial messenger RNA)
RN  - EC 2.7.7.- (RNA Nucleotidyltransferases)
RN  - EC 2.7.7.- (UTP-RNA uridylyltransferase)
RN  - UT0S826Z60 (Uridine Triphosphate)
SB  - IM
MH  - Binding Sites
MH  - Mutagenesis
MH  - Protein Structure, Tertiary
MH  - Protozoan Proteins/*chemistry/genetics/metabolism
MH  - *RNA Editing
MH  - RNA Nucleotidyltransferases/*chemistry/metabolism
MH  - RNA, Messenger/chemistry/*metabolism
MH  - RNA, Mitochondrial
MH  - RNA, Protozoan/chemistry/*metabolism
MH  - Trypanosoma/*enzymology/genetics
MH  - Uridine Triphosphate/metabolism
PMC - PMC2885561
MID - NIHMS200237
COIS- This manuscript has been approved by all authors. Authors declare no conflicts of 
      interest or competing financial interests.
EDAT- 2010/04/27 06:00
MHDA- 2010/09/09 06:00
PMCR- 2011/06/25
CRDT- 2010/04/27 06:00
PHST- 2009/12/09 00:00 [received]
PHST- 2010/04/14 00:00 [revised]
PHST- 2010/04/19 00:00 [accepted]
PHST- 2010/04/27 06:00 [entrez]
PHST- 2010/04/27 06:00 [pubmed]
PHST- 2010/09/09 06:00 [medline]
PHST- 2011/06/25 00:00 [pmc-release]
AID - S0022-2836(10)00409-2 [pii]
AID - 10.1016/j.jmb.2010.04.035 [doi]
PST - ppublish
SO  - J Mol Biol. 2010 Jun 25;399(5):696-706. doi: 10.1016/j.jmb.2010.04.035. Epub 2010 
      Apr 24.