Establishment of a Novel in Vivo Sex-specific Splicing Assay System to Identify a Trans-acting Factor That Negatively Regulates Splicing of Bombyx Mori Dsx Female Exons

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 2007 Oct 31

PMID 17967886

Citations 34

Authors

Masataka G Suzuki

Shigeo Imanishi

Naoshi Dohmae

Tomoe Nishimura

Toru Shimada

Shogo Matsumoto

Affiliations

Soon will be listed here.

Abstract

The Bombyx mori homolog of doublesex, Bmdsx, plays an essential role in silkworm sexual development. Exons 3 and 4 of Bmdsx pre-mRNA are specifically excluded in males. To explore how this occurs, we developed a novel in vivo sex-specific splicing assay system using sexually differentiated cultured cells. A series of mutation analyses using a Bmdsx minigene with this in vivo splicing assay system identified three distinct sequences (CE1, CE2, and CE3) positioned in exon 4 as exonic splicing silencers responsible for male-specific splicing. Gel shift analysis showed that CE1 binds to a nuclear protein from male cells but not that from female cells. Mutation of UAA repeats within CE1 inhibited the binding of the nuclear protein to the RNA and caused female-specific splicing in male cells. We have identified BmPSI, a Bombyx homolog of P-element somatic inhibitor (PSI), as the nuclear factor that specifically binds CE1. Down-regulation of endogenous BmPSI by RNA interference significantly increased female-specific splicing in male cells. This is the first report of a PSI homolog implicated in the regulated sex-specific splicing of dsx pre-mRNA.

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