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PMID: 16882972 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Specific association of Piwi with rasiRNAs derived from retrotransposon and heterochromatic regions in the Drosophila genome.

Genes & development ·Vol. 20 ·No. 16 ·2006-08-15 ·Pages 2214-22

Saito K, Nishida KM, Mori T, Kawamura Y, Miyoshi K, Nagami T, Siomi H, Siomi MC

Abstract

In Drosophila, Piwi (P-element-induced wimpy testis), which encodes a protein of the Argonaute family, is essential for germ stem cell self-renewal. Piwi has recently been shown to be a nuclear protein involved in gene silencing of retrotransposons and controlling their mobilization in the male germline. However, little is known about the molecular mechanisms of Piwi-dependent gene silencing. Here we show that endogenous Piwi immunopurified from ovary specifically associates with small RNAs of 25-29 nucleotides in length. Piwi-associated small RNAs were identified by cloning and sequencing as repeat-associated small interfering RNAs (rasiRNAs) derived from repetitive regions, such as retrotransposon and heterochromatic regions, in the Drosophila genome. Northern blot analyses revealed that in vivo Piwi does not associate with microRNAs (miRNAs) and that guide siRNA was not loaded onto Piwi when siRNA duplex was added to ovary lysate. In vitro, recombinant Piwi exhibits target RNA cleavage activity. These data together imply that Piwi functions in nuclear RNA silencing as Slicer by associating specifically with rasiRNAs originating from repetitive targets.

MeSH Terms
Animals Argonaute Proteins Blotting, Northern Drosophila Proteins Drosophila melanogaster/genetics,metabolism Female Genome, Insect Heterochromatin/genetics Male MicroRNAs/metabolism Models, Genetic Molecular Sequence Data Ovary/metabolism Proteins/immunology,metabolism RNA Interference RNA, Small Interfering/metabolism RNA-Induced Silencing Complex Repetitive Sequences, Nucleic Acid Retroelements/genetics Testis/metabolism
Chemicals
Argonaute Proteins Drosophila Proteins Heterochromatin MicroRNAs Proteins RNA, Small Interfering RNA-Induced Silencing Complex Retroelements piwi protein, Drosophila
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Saito Kuniaki
Institute for Genome Research, University of Tokushima, Tokushima 770-8503, Japan.
Nishida Kazumichi M
Mori Tomoko
Kawamura Yoshinori
Miyoshi Keita
Nagami Tomoko
Siomi Haruhiko
Siomi Mikiko C
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1454806
Published
2006-08-15
Epub
2006-00-01
Pages
2214-22
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC1553205
Subset
IM
Databases
GENBANK
AF237761, AF364549, AF364550, AF365402, AF420242, AF456125, AF492764, AF533772, AJ132547, AJ278684, AJ487856, AJ542581, AJ635224, AY180916, AY180917, AY180918, AY183918, AY183919, AY600955, AY928610, M12927, U06920, U14101, U23420, X03431, X04132, X04456, X06950, X07656, X14037, X51967, X51968, X59545, X77571, X95908, X98424, Z31368
External Links
PubMed source DOI: 10.1101/gad.1454806 PMC Full Text View journal details
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