Blanks, a nuclear siRNA/dsRNA-binding complex component, is required for Drosophila spermiogenesis

Small RNAs and a diverse array of protein partners control gene expression in eukaryotes through a variety of mechanisms. By combining siRNA affinity chromatography and mass spectrometry, we have identified the double-stranded RNA-binding domain protein Blanks to be an siRNA- and dsRNA-binding prote...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2011-02, Vol.108 (8), p.3204-3209
Main Authors: Gerbasi, Vincent R, Preall, Jonathan B, Golden, Daniel E, Powell, David W, Cummins, Timothy D, Sontheimer, Erik J
Format: Article
Language:English
Subjects:
Affinity chromatography
Animals
Biological Sciences
Cell lines
Chromatography
Drosophila
Drosophila - physiology
Gene expression
Gene Expression Regulation - physiology
Germ cells
Immunity, Innate - genetics
Individualization
Insects
Male
Mass spectrometry
Messenger RNA
Proteins
Ribonucleic acid
RNA
RNA Interference
RNA, Double-Stranded
RNA, Small Interfering
RNA-Binding Proteins - physiology
Small interfering RNA
Spermatogenesis
Spermatozoa
Stem cells
Testes
Testis - chemistry
Testis - metabolism
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Summary:Small RNAs and a diverse array of protein partners control gene expression in eukaryotes through a variety of mechanisms. By combining siRNA affinity chromatography and mass spectrometry, we have identified the double-stranded RNA-binding domain protein Blanks to be an siRNA- and dsRNA-binding protein from Drosophila S2 cells. We find that Blanks is a nuclear factor that contributes to the efficiency of RNAi. Biochemical fractionation of a Blanks-containing complex shows that the Blanks complex is unlike previously described RNA-induced silencing complexes and associates with the DEAD-box helicase RM62, a protein previously implicated in RNA silencing. In flies, Blanks is highly expressed in testes tissues and is necessary for postmeiotic spermiogenesis, but loss of Blanks is not accompanied by detectable transposon derepression. Instead, genes related to innate immunity pathways are up-regulated in blanks mutant testes. These results reveal Blanks to be a unique component of a nuclear siRNA/dsRNA-binding complex that contributes to essential RNA silencing-related pathways in the male germ line.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1009781108