Proteomics identification of Drosophila small interfering RNA-associated factors.
The Drosophila melanogaster RNA-induced silencing complex (RISC) forms a large ribonucleoprotein particle on small interfering RNAs (siRNAs) and catalyzes target mRNA cleavage during RNA interference (RNAi). Dicer-2, R2D2, Loquacious, and Argonaute-2 are examples of RISC-associated factors that are involved in RNAi. Holo-RISC is an approximately 80 S small interfering ribonucleoprotein, which suggests that there are many additional proteins that participate in the RNAi pathway. In this study, we used siRNA affinity capture combined with mass spectrometry to identify novel components of the Drosophila RNAi machinery. Our study identified both established RISC components and novel siRNA-associated factors, many of which contain domains that are consistent with potential roles in RNAi. Functional analysis of these novel siRNA-associated proteins suggests that these factors may play an important role in RNAi.
Gerbasi
Vincent R
VR
Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, Illinois 60208-3500, USA.
Golden
Daniel E
DE
Hurtado
Sara B
SB
Sontheimer
Erik J
EJ
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D016000:Cluster Analysis; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster; D040901:Proteomics; D034741:RNA, Small Interfering / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
D029721:Drosophila Proteins; D034741:RNA, Small Interfering
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