Functional interaction between the carboxy-terminal domain of RNA polymerase II and pre-mRNA splicing

被引:121
作者
Du, L
Warren, SL
机构
[1] YALE UNIV, SCH MED, DEPT PATHOL, BRADY MEM LABS, NEW HAVEN, CT 06520 USA
[2] YALE UNIV, SCH MED, DEPT GENET, NEW HAVEN, CT 06520 USA
关键词
D O I
10.1083/jcb.136.1.5
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
In the preceding study we found that Sm snRNPs and SerArg (SR) family proteins co-immunoprecipitate with Pol II molecules containing a hyperphosphorylated CTD (Kim et al., 1997). The association between Pol IIo and splicing factors is maintained in the absence of pre-mRNA, and the polymerase need not be transcriptionally engaged (Kim et al., 1997). The latter findings led us to hypothesize that a phosphorylated form of the CTD interacts with pre-mRNA splicing components in vivo. To test this idea, a nested set of CTD-derived proteins was assayed for the ability to alter the nuclear distribution of splicing factors, and to interfere with splicing in vivo. Proteins containing heptapeptides 1-52 (CTD52), 1-32 (CTD32), 1-26 (CTD26), 1-13 (CTD13), 1-6 (CTD6), 1-3 (CTD3), or 1 (CTD1) were expressed in mammalian cells. The CTD-derived proteins become phosphorylated in vivo, and accumulate in the nucleus even though they lack a conventional nuclear localization signal. CTD52 induces a selective reorganization of splicing factors from discrete nuclear domains to the diffuse nucleoplasm, and significantly, it blocks the accumulation of spliced, but not unspliced, human P-globin transcripts. The extent of splicing factor disruption, and the degree of inhibition of splicing, are proportional to the number of heptapeptides added to the protein. The above results indicate a functional interaction between Pol II's CTD and pre-mRNA splicing.
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页码:5 / 18
页数:14
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