Sex-lethal interacts with splicing factors in vitro and in vivo.
The Drosophila sex determination gene Sex-lethal controls its own expression and the expression of downstream target genes such as transformer by regulating RNA splicing. Genetic and molecular studies have established that Sxl requires the product of another gene, snf, to autoregulate the splicing of its own transcripts. snf has recently been shown to encode a Drosophila U1 and U2 small nuclear ribonucleoprotein particle protein. In the work reported here, we demonstrate that the Sxl and Snf proteins can interact directly in vitro and that these two proteins are part of an RNase-sensitive complex in vivo which can be immunoprecipitated with the Sxl antibody. Unlike bulk Snf protein, which sediments slowly in sucrose gradients, the Snf protein associated with Sxl is in a large, rapidly sedimenting complex. Detailed characterization of the Sxl-Snf complexes from cross-linked extracts indicates that these complexes contain additional small nuclear ribonucleoprotein particle proteins and the U1 and U2 small nuclear RNAs. Finally, consistent with the RNase sensitivity of the Sxl-Snf complexes, Sxl transcripts can also be immunoprecipitated by Sxl antibodies. On the basis of the physical interactions between Sxl and Snf, we present a model for Sxl splicing regulation. This model helps explain how the Sxl protein is able to promote the sex-specific splicing of Sxl transcripts, utilizing target sequences that are distant from the regulated splice sites.
Deshpande
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Department of Molecular Biology, Princeton University, New Jersey 08544, USA.
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics*; D005786:Gene Expression Regulation; D046911:Macromolecular Substances; D008957:Models, Genetic; D008969:Molecular Sequence Data; D012326:RNA Splicing / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000276:immunology / Q000502:physiology*; D017412:Ribonucleoprotein, U1 Small Nuclear / Q000502:physiology*; D012732:Sex Determination Analysis
D000911:Antibodies, Monoclonal; D029721:Drosophila Proteins; D046911:Macromolecular Substances; D016601:RNA-Binding Proteins; D017412:Ribonucleoprotein, U1 Small Nuclear; C067777:Sxl protein, Drosophila; C089969:snf protein, Drosophila