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Characterization of the piRNA Complex from Rat Testes
Nelson C. Lau,1*Anita G. Seto,1*Jinkuk Kim,2,3Satomi Kuramochi-Miyagawa,4Toru Nakano,4David P. Bartel,3,5Robert E. Kingston1
Small noncoding RNAs regulate processes essential for cell growthand development, including mRNA degradation, translational repression,and transcriptional gene silencing (TGS). During a search forcandidate mammalian factors for TGS, we purified a complex thatcontains small RNAs and Riwi, the rat homolog to human Piwi.The RNAs, frequently 29 to 30 nucleotides in length, are calledPiwi-interacting RNAs (piRNAs), 94% of which map to 100 defined(101 kb) genomic regions. Within these regions, the piRNAs generallydistribute across only one genomic strand or distribute on twostrands but in a divergent, nonoverlapping manner. Preparationsof piRNA complex (piRC) contain rRecQ1, which is homologousto qde-3 from Neurospora, a gene implicated in silencing pathways.Piwi has been genetically linked to TGS in flies, and sliceractivity cofractionates with the purified complex. These resultsare consistent with a gene-silencing role for piRC in mammals.
1 Department of Molecular Biology, Massachusetts General Hospital, 185 Cambridge Street, Boston, MA 02114, USA. 2 Harvard-MIT Division of Health Sciences and Technology, E18-435, 77 Massachusetts Avenue, Cambridge, MA 02139, USA. 3 Howard Hughes Medical Institute and Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, MA 02142, USA. 4 Department of Molecular Cell Biology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita-shi, Osaka 565-0871, Japan. 5 Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
* These authors contributed equally to this work.
To whom correspondence should be addressed. E-mail: kingston{at}molbio.mgh.harvard.edu
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