Dataset Information

Proteins that interact physically with Drosophila blanks

ABSTRACT: RNA interference targets aberrant transcripts with cognate small interfering RNAs, which derive from double-stranded RNA precursors through nucleolytic processing. Several functional screens have identified Drosophila blanks/lump (CG10630) as a facilitator of RNAi in cultured cells, yet its molecular function has remained unknown. The protein carries two dsRNA binding domains (dsRBD) and blanks mutant males have a spermatogenesis defect. We demonstrate that blanks selectively boosts RNAi triggered by dsRNA of nuclear origin. Blanks binds dsRNA in vitro, shuttles between nucleus and cytoplasm and the abundance of certain endogenous siRNAs is reduced in blanks mutant testes. Transgenic expression of a Blanks protein variant with increased nuclear retention reduces endogenous siRNA abundance unless the protein also carries a mutation that prevents binding of dsRNA to the second dsRBD. Blanks thus facilitates the export of dsRNA to the cytoplasm for further processing by the canonical RNAi machinery. In contrast, rescue of male fertility in blanks mutant animals was independent of RNA binding to the second dsRBD, consistent with a previous report that linked fertility to the first dsRBD of Blanks. The role of blanks in spermatogenesis appears thus unrelated to its role in dsRNA export.

INSTRUMENT(S): LTQ Orbitrap XL

ORGANISM(S): Drosophila Melanogaster (fruit Fly)

TISSUE(S): Cell Culture

SUBMITTER: Klaus Förstemann

LAB HEAD: Klaus Förstemann

PROVIDER: PXD013792 | Pride | 2020-05-26

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	Kunzelmann_10_20170126.raw	Raw
	Kunzelmann_11_20170126.raw	Raw
	Kunzelmann_12_20170126.raw	Raw
	Kunzelmann_1_20170126.raw	Raw
	Kunzelmann_2_20170126.raw	Raw

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Publications

Trafficking of siRNA precursors by the dsRBD protein Blanks in Drosophila.

Nitschko Volker V Kunzelmann Stefan S Fröhlich Thomas T Arnold Georg J GJ Förstemann Klaus K

Nucleic acids research 20200401 7

RNA interference targets aberrant transcripts with cognate small interfering RNAs, which derive from double-stranded RNA precursors. Several functional screens have identified Drosophila blanks/lump (CG10630) as a facilitator of RNAi, yet its molecular function has remained unknown. The protein carries two dsRNA binding domains (dsRBD) and blanks mutant males have a spermatogenesis defect. We demonstrate that blanks selectively boosts RNAi triggered by dsRNA of nuclear origin. Blanks binds dsRNA ...[more]

PMID: 32025726

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