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Science 8 June 1990:
Vol. 248. no. 4960, pp. 1200 - 1208
DOI: 10.1126/science.2161558
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Articles

Science, Vol 248, Issue 4960, 1200-1208
Copyright © 1990 by American Association for the Advancement of Science

articles

Regulated splicing of the Drosophila P transposable element third intron in vitro: somatic repression

CW Siebel and DC Rio

Whitehead Institute for Biomedical Research, Nine Cambridge Center, MA 02142.

In eukaryotic cells alternative splicing of messenger RNA precursors (pre-mRNA's) is a means of regulating gene expression. Although a number of the components that participate in regulating some alternative splicing events have been identified by molecular genetic procedures, the elucidation of the biochemical mechanisms governing alternative splicing requires in vitro reaction systems. The tissue specificity of P element transposition in Drosophila depends on the germline restriction of pre-mRNA splicing of the P element third intron (IVS3). Drosophila P element IVS3 pre-mRNA substrates were spliced accurately in vitro in heterologous human cell extracts but not in Drosophila somatic cell splicing extracts. Components in Drosophila somatic cell extracts that specifically inhibited IVS3 splicing in vitro were detected by a complementation assay. Biochemical assays for Drosophila RNA binding proteins were then used to detect a 97-kilodalton protein that interacts specifically with 5' exon sequences previously implicated in the control of IVS3 splicing in vivo. Inhibition of IVS3 splicing in vitro could be correlated with binding of the 97-kD protein to 5' exon sequences, suggesting that one aspect of IVS3 tissue-specific splicing involves somatic repression by specific RNA-protein interactions.


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U1 snRNP can influence 3'-splice site selection as well as 5'-splice site selection..
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