Publication Date:
2011-07-15
Description:
A hallmark of RNA silencing is a class of approximately 22-nucleotide RNAs that are processed from double-stranded RNA precursors by Dicer. Accurate processing by Dicer is crucial for the functionality of microRNAs (miRNAs). The current model posits that Dicer selects cleavage sites by measuring a set distance from the 3' overhang of the double-stranded RNA terminus. Here we report that human Dicer anchors not only the 3' end but also the 5' end, with the cleavage site determined mainly by the distance ( approximately 22 nucleotides) from the 5' end (5' counting rule). This cleavage requires a 5'-terminal phosphate group. Further, we identify a novel basic motif (5' pocket) in human Dicer that recognizes the 5'-phosphorylated end. The 5' counting rule and the 5' anchoring residues are conserved in Drosophila Dicer-1, but not in Giardia Dicer. Mutations in the 5' pocket reduce processing efficiency and alter cleavage sites in vitro. Consistently, miRNA biogenesis is perturbed in vivo when Dicer-null embryonic stem cells are replenished with the 5'-pocket mutant. Thus, 5'-end recognition by Dicer is important for precise and effective biogenesis of miRNAs. Insights from this study should also afford practical benefits to the design of small hairpin RNAs.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4693635/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4693635/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Park, Jong-Eun -- Heo, Inha -- Tian, Yuan -- Simanshu, Dhirendra K -- Chang, Hyeshik -- Jee, David -- Patel, Dinshaw J -- Kim, V Narry -- P30 CA008748/CA/NCI NIH HHS/ -- R01 AI068776/AI/NIAID NIH HHS/ -- England -- Nature. 2011 Jul 13;475(7355):201-5. doi: 10.1038/nature10198.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉School of Biological Sciences, Seoul National University, Seoul 151-742, Korea.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/21753850" target="_blank"〉PubMed〈/a〉
Keywords:
Amino Acid Sequence
;
Animals
;
Base Sequence
;
Binding Sites/genetics
;
DEAD-box RNA Helicases/deficiency/genetics/*metabolism
;
Drosophila Proteins/metabolism
;
Embryonic Stem Cells/metabolism
;
Evolution, Molecular
;
Giardia/enzymology
;
HEK293 Cells
;
Humans
;
MicroRNAs/biosynthesis/chemistry/genetics/*metabolism
;
Molecular Sequence Data
;
Mutant Proteins/chemistry/genetics/metabolism
;
Mutation/genetics
;
Phosphates/metabolism
;
Phosphorylation
;
RNA Helicases/metabolism
;
Ribonuclease III/deficiency/genetics/*metabolism
;
Substrate Specificity/genetics
Print ISSN:
0028-0836
Electronic ISSN:
1476-4687
Topics:
Biology
,
Chemistry and Pharmacology
,
Medicine
,
Natural Sciences in General
,
Physics
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