» Articles » PMID: 17967886

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
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.

Citing Articles

Cell-based analysis reveals that sex-determining gene signals in are pivotally changed by male-killing .

Herran B, Sugimoto T, Watanabe K, Imanishi S, Tsuchida T, Matsuo T PNAS Nexus. 2023; 2(1):pgac293.

PMID: 36712932 PMC: 9837667. DOI: 10.1093/pnasnexus/pgac293.


A database of crop pest cell lines.

Arya S, Goodman C, Stanley D, Palli S In Vitro Cell Dev Biol Anim. 2022; 58(8):719-757.

PMID: 35994130 DOI: 10.1007/s11626-022-00710-w.


Theme and variation in the evolution of insect sex determination.

Laslo M, Just J, Angelini D J Exp Zool B Mol Dev Evol. 2022; 340(2):162-181.

PMID: 35239250 PMC: 10078687. DOI: 10.1002/jez.b.23125.


Identification and Characterization of the Masculinizing Function of the Gene.

Deng Z, Zhang Y, Li Y, Huang K, Chen X, Zhang M Int J Mol Sci. 2021; 22(16).

PMID: 34445352 PMC: 8395511. DOI: 10.3390/ijms22168650.


The Sex Determination Cascade in the Silkworm.

Yang X, Chen K, Wang Y, Yang D, Huang Y Genes (Basel). 2021; 12(2).

PMID: 33672402 PMC: 7926724. DOI: 10.3390/genes12020315.


References
1.
Kan J, Green M . Pre-mRNA splicing of IgM exons M1 and M2 is directed by a juxtaposed splicing enhancer and inhibitor. Genes Dev. 1999; 13(4):462-71. PMC: 316470. DOI: 10.1101/gad.13.4.462. View

2.
Tian M, Maniatis T . A splicing enhancer exhibits both constitutive and regulated activities. Genes Dev. 1994; 8(14):1703-12. DOI: 10.1101/gad.8.14.1703. View

3.
Zhu L, Wilken J, Phillips N, Narendra U, Chan G, Stratton S . Sexual dimorphism in diverse metazoans is regulated by a novel class of intertwined zinc fingers. Genes Dev. 2000; 14(14):1750-64. PMC: 316782. View

4.
Siebel C, Fresco L, Rio D . The mechanism of somatic inhibition of Drosophila P-element pre-mRNA splicing: multiprotein complexes at an exon pseudo-5' splice site control U1 snRNP binding. Genes Dev. 1992; 6(8):1386-401. DOI: 10.1101/gad.6.8.1386. View

5.
Mougin A, Gregoire A, Banroques J, Segault V, Fournier R, Brule F . Secondary structure of the yeast Saccharomyces cerevisiae pre-U3A snoRNA and its implication for splicing efficiency. RNA. 1996; 2(11):1079-93. PMC: 1369438. View