PMID- 19264675
OWN - NLM
STAT- MEDLINE
DCOM- 20090508
LR  - 20211020
IS  - 1469-9001 (Electronic)
IS  - 1355-8382 (Print)
IS  - 1355-8382 (Linking)
VI  - 15
IP  - 5
DP  - 2009 May
TI  - E. coli RNase III(E38A) generates discrete-sized products from long dsRNA.
PG  - 984-91
LID - 10.1261/rna.1196509 [doi]
AB  - Ribonuclease III (RNase III) represents a highly conserved family of 
      double-strand-specific endoribonucleases that are important for RNA processing 
      and post-transcriptional gene regulation in both prokaryotes and eukaryotes. We 
      constructed a single amino acid substitution (E38A) of RNase III that shows a 
      unique and useful enzymatic activity. It produces a dsRNA product of a discrete 
      size migrating as 23 base pairs (bp) when given a long dsRNA as a substrate in an 
      easy-to-control reaction. We demonstrate that the RNase III(E38A) mutant produces 
      the 23-bp dsRNA product by making a double-strand cleavage of the long dsRNA 
      substrate with the product being protected from further digestion. Using the 
      hairpin RNA R1.1 as a substrate, RNase III(E38A) cleaves at the primary site and 
      remains bound to the RNA, thereby preventing cleavage at the secondary site. The 
      23-bp dsRNA product is demonstrated to be a pool of dsRNAs representative of the 
      long dsRNA substrate and has RNA interference activity in mammalian tissue 
      culture transfection experiments. The RNA interference activity suggests that the 
      23-bp dsRNA product has typical 2-nucleotide 3' overhangs and behaves as siRNA 
      thereby making it a useful tool in RNA interference experiments.
FAU - Xiao, Jianping
AU  - Xiao J
AD  - New England Biolabs, Ipswich, Massachusetts 01938, USA.
FAU - Feehery, Caitlin E
AU  - Feehery CE
FAU - Tzertzinis, George
AU  - Tzertzinis G
FAU - Maina, Claude V
AU  - Maina CV
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090304
PL  - United States
TA  - RNA
JT  - RNA (New York, N.Y.)
JID - 9509184
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (RNA, Double-Stranded)
RN  - EC 3.1.26.3 (Ribonuclease III)
RN  - EC 3.1.26.3 (ribonuclease III, E coli)
SB  - IM
MH  - Amino Acid Substitution
MH  - Base Sequence
MH  - Escherichia coli Proteins/*genetics/*metabolism
MH  - Nucleic Acid Conformation
MH  - RNA Interference
MH  - RNA, Double-Stranded/chemistry/*metabolism
MH  - Ribonuclease III/*genetics/*metabolism
PMC - PMC2673077
EDAT- 2009/03/07 09:00
MHDA- 2009/05/09 09:00
PMCR- 2009/11/01
CRDT- 2009/03/07 09:00
PHST- 2009/03/07 09:00 [entrez]
PHST- 2009/03/07 09:00 [pubmed]
PHST- 2009/05/09 09:00 [medline]
PHST- 2009/11/01 00:00 [pmc-release]
AID - rna.1196509 [pii]
AID - RA [pii]
AID - 10.1261/rna.1196509 [doi]
PST - ppublish
SO  - RNA. 2009 May;15(5):984-91. doi: 10.1261/rna.1196509. Epub 2009 Mar 4.