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A complex containing CstF-64 and the SL2 ... s-splicing in C elegans operons
A complex containing CstF-64 and the SL2 snRNP connects mRNA 3' end formation and trans-splicing in C. elegans operons.
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Polycistronic pre-mRNAs from Caenorhabditis elegans are processed by 3' end formation of the upstream mRNA and SL2-specific trans-splicing of the downstream mRNA. These processes usually occur within an approximately 100-nucleotide region and are mechanistically coupled. In this paper, we report a complex in C. elegans extracts containing the 3' end formation protein CstF-64 and the SL2 snRNP. This complex, immunoprecipitated with alphaCstF-64 antibody, contains SL2 RNA, but not SL1 RNA or other U snRNAs. Using mutational analysis we have been able to uncouple SL2 snRNP function and identity. SL2 RNA with a mutation in stem/loop III is functional in vivo as a trans-splice donor, but fails to splice to SL2-accepting trans-splice sites, suggesting that it has lost its identity as an SL2 snRNP. Importantly, stem/loop III mutations prevent association of SL2 RNA with CstF-64. In contrast, a mutation in stem II that inactivates the SL2 snRNP still permits complex formation with CstF-64. Therefore, SL2 RNA stem/loop III is required for both SL2 identity and formation of a complex containing CstF-64, but not for trans-splicing. These results provide a molecular framework for the coupling of 3' end formation and trans-splicing in the processing of polycistronic pre-mRNAs from C. elegans operons.
Evans D, Perez I, MacMorris M, Leake D, Wilusz CJ, Blumenthal T
Genes & development
2001-10-01 00:00
15
19
2562-71
Animals,Base Sequence,Caenorhabditis elegans,DNA Primers,Mutation,Operon,RNA Splicing,RNA, Messenger,RNA, Spliced Leader,RNA-Binding Proteins,Ribonucleoproteins, Small Nuclear,mRNA Cleavage and Polyadenylation Factors,DNA Primers,RNA, Messenger,RNA, Spliced Leader,RNA-Binding Proteins,Ribonucleoproteins, Small Nuclear,mRNA Cleavage and Polyadenylation Factors
Department of Biochemistry and Molecular Genetics, University of Colorado School of Medicine, Denver, CO 80262, USA
Genes Dev.
NIGMS GM 42432
0890-9369
10.1101/gad.920501
1
True
11581161
