Calvo Olga, Manley James L
Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
EMBO J. 2005 Mar 9;24(5):1009-20. doi: 10.1038/sj.emboj.7600575. Epub 2005 Feb 3.
Transcription and processing of mRNA precursors are coordinated events that require numerous complex interactions to ensure that they are successfully executed. We described previously an unexpected association between a transcription factor, PC4 (or Sub1 in yeast), and an mRNA polyadenylation factor, CstF-64 (Rna15 in yeast), and provided evidence that this was important for efficient transcription elongation. Here we provide insight into the mechanism by which this occurs. We show that Sub1 and Rna15 are recruited to promoters and present along the length of several yeast genes. Allele-specific genetic interactions between SUB1 and genes encoding an RNA polymerase II (RNAP II)-specific kinase (KIN28) and phosphatase (FCP1) suggest that Sub1 influences and/or is sensitive to the phosphorylation status of elongating RNAP II. Remarkably, we find that cells lacking Sub1 display decreased accumulation of Fcp1, altered RNAP II phosphorylation and decreased crosslinking of RNAP II to transcribed genes. Our data provide evidence that Rna15 and Sub1 are present along the length of several genes and that Sub1 facilitates elongation by influencing enzymes that modify RNAP II.
mRNA前体的转录和加工是相互协调的过程,需要众多复杂的相互作用以确保它们能成功进行。我们之前描述了一种转录因子PC4(酵母中的Sub1)与一种mRNA聚腺苷酸化因子CstF - 64(酵母中的Rna15)之间意外的关联,并提供证据表明这对高效转录延伸很重要。在此我们深入探讨了其发生机制。我们发现Sub1和Rna15被招募到启动子区域,并存在于多个酵母基因的全长范围内。SUB1与编码RNA聚合酶II(RNAP II)特异性激酶(KIN28)和磷酸酶(FCP1)的基因之间的等位基因特异性遗传相互作用表明,Sub1影响延伸中的RNAP II的磷酸化状态和/或对其敏感。值得注意的是,我们发现缺乏Sub1的细胞中Fcp1的积累减少,RNAP II磷酸化改变,且RNAP II与转录基因的交联减少。我们的数据提供了证据,表明Rna15和Sub1存在于多个基因的全长范围内且Sub1通过影响修饰RNAP II的酶来促进转录延伸。