A role for CF1A 3' end processing complex in promoter-associated transcription.
A role for CF1A 3' end processing complex in promoter-associated transcription.
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DOI：
10.1371/journal.pgen.1003722
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发表时间：
2013
期刊：
影响因子：
4.5
通讯作者：
Ansari A
中科院分区：
文献类型：
--
作者：
Al Husini N;Kudla P;Ansari A
The Cleavage Factor 1A (CF1A) complex, which is required for the termination of transcription in budding yeast, occupies the 3′ end of transcriptionally active genes. We recently demonstrated that CF1A subunits also crosslink to the 5′ end of genes during transcription. The presence of CF1A complex at the promoter suggested its possible involvement in the initiation/reinitiation of transcription. To check this possibility, we performed transcription run-on assay, RNAP II-density ChIP and strand-specific RT-PCR analysis in a mutant of CF1A subunit Clp1. As expected, RNAP II read through the termination signal in the temperature-sensitive mutant of clp1 at elevated temperature. The transcription readthrough phenotype was accompanied by a decrease in the density of RNAP II in the vicinity of the promoter region. With the exception of TFIIB and TFIIF, the recruitment of the general transcription factors onto the promoter, however, remained unaffected in the clp1 mutant. These results suggest that the CF1A complex affects the recruitment of RNAP II onto the promoter for reinitiation of transcription. Simultaneously, an increase in synthesis of promoter-initiated divergent antisense transcript was observed in the clp1 mutant, thereby implicating CF1A complex in providing directionality to the promoter-bound polymerase. Chromosome Conformation Capture (3C) analysis revealed a physical interaction of the promoter and terminator regions of a gene in the presence of a functional CF1A complex. Gene looping was completely abolished in the clp1 mutant. On the basis of these results, we propose that the CF1A-dependent recruitment of RNAP II onto the promoter for reinitiation and the regulation of directionality of promoter-associated transcription are accomplished through gene looping. The termination of transcription requires two major multisubunit complexes in budding yeast. These termination complexes are localized at the 3′ end of genes. Recent studies have found the termination factors occupying the 5′ end of genes as well. In this study, we investigate the physiological role of a termination factor at the 5′ end of a gene. Our results show that the CF1 termination complex affects the recruitment of the transcription enzyme RNAP II onto the promoter for reinitiation of transcription. The complex also affects the directionality of transcription of the promoter-bound polymerase. We also found that the looped gene conformation was disrupted in the absence of a functional termination complex. The overall conclusion of these results is that the terminator-bound factors contact the 5′ end of genes due to gene looping, and affect both the recruitment of the polymerase at the promoter for reinitiation, and directionality of the promoter-initiated transcription. Thus, the role of termination factors is not restricted to the 3′ end of the gene, but they are also involved in promoter-associated transcription.
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