Key result
Selective cleavage of U4 or U6 RNA inhibits pre-mRNA splicing in HeLa cell extracts.
Demonstrates that intact U4/U6 small nuclear ribonucleoprotein is required for pre-mRNA splicing in vitro.
No immediate clinical implications from in vitro data; leaves open validation in disease models and therapeutic development.
Selective cleavage of U4 or U6 RNA in a HeLa cell nuclear extract inhibits splicing of pre-mRNAs containing an adenovirus or a simian virus 40 intron. RNAs in the U4/U6 small nuclear ribonucleoprotein (snRNP) were specifically degraded with RNAase H and deoxyoligonucleotides. Two oligomers complementary to U4 RNA and two complementary to U6 RNA cleave their target RNAs and inhibit the appearance of both spliced products and reaction intermediates. Splicing is reconstituted by mixing an extract containing cleaved U4 or U6 RNA with one in which splicing has been inhibited by degrading U2 RNA. All four abundant snRNPs, containing U1, U2, U5, or U4 and U6 RNAs, are now implicated in pre-mRNA splicing. Possible interactions of the U4/U6 snRNP with other components of the splicing complex are discussed.
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Black et al. (1986) studied this question. Selective cleavage of U4 or U6 RNA was evaluated on splicing of pre-mRNAs containing an adenovirus or a simian virus 40 intron. Selective cleavage of U4 or U6 RNA in a HeLa cell nuclear extract inhibits splicing of pre-mRNAs, implicating U4 and U6 RNAs in pre-mRNA splicing.
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