Role for PSF in mediating transcriptional activator-dependent stimulation of pre-mRNA processing in vivo

被引:101
作者
Rosonina, E
Ip, JYY
Calarco, JA
Bakowski, MA
Emili, A
McCracken, S
Tucker, P
Ingles, CJ
Blencowe, BJ
机构
[1] Univ Toronto, Charles H Best Inst, Banting & Best Dept Med Res, Toronto, ON M5G 1L6, Canada
[2] Univ Toronto, Dept Mol & Med Genet, Toronto, ON M5G 1L6, Canada
[3] Univ Toronto, Proteom & Bioinformat Program, Toronto, ON M5G 1L6, Canada
[4] Univ Texas, Inst Mol & Cellular Biol, Sect Mol Genet & Microbiol, Austin, TX 78712 USA
关键词
D O I
10.1128/MCB.25.15.6734-6746.2005
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In a recent study, we provided evidence that strong promoter-bound transcriptional activators result in higher levels of splicing and 3'-end cleavage of nascent pre-mRNA than do weak promoter-bound activators and that this effect of strong activators requires the carboxyl-terminal domain (CTD) of RNA polymerase 11 (pol II). In the present study, we have investigated the mechanism of activator- and CTD-mediated stimulation of pre-mRNA processing. Affinity chromatography experiments reveal that two factors previously implicated in the coupling of transcription and pre-mRNA processing, PSF and p54(nrb)/NonO, preferentially bind a strong rather than a weak activation domain. Elevated expression in human 293 cells of PSF bypasses the requirement for a strong activator to promote efficient splicing and Y-end cleavage. Truncation of the pol 11 CTD, which consists of 52 repeats of the consensus heptapeptide sequence YSPTSPS, to 15 heptapeptide repeats prevents PSF-dependent stimulation of splicing and Y-end cleavage. Moreover, PSF and p54(nrb)/NonO bind in vitro to the wild-type CTD but not to the truncated 15-repeat CTD, and domains in PSF that are required for binding to activators and to the CTD are also important for the stimulation of pre-mRNA processing. Interestingly, activator- and CTD-dependent stimulation of splicing mediated by PSF appears to primarily affect the removal of first introns. Collectively, these results suggest that the recruitment of PSF to activated promoters and the pol II CTD provides a mechanism by which transcription and pre-mRNA processing are coordinated within the cell.
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收藏
页码:6734 / 6746
页数:13
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