Uhler Jay P, Hertel Christina, Svejstrup Jesper Q
Mechanisms of Transcription Laboratory, Cancer Research UK London Research Institute, Clare Hall Laboratories, South Mimms EN6 3LD, United Kingdom.
Proc Natl Acad Sci U S A. 2007 May 8;104(19):8011-6. doi: 10.1073/pnas.0702431104. Epub 2007 Apr 30.
Noncoding, or intergenic, transcription by RNA polymerase II (RNAPII) is remarkably widespread in eukaryotic organisms, but the effects of such transcription remain poorly understood. Here we show that noncoding transcription plays a role in activation, but not repression, of the Saccharomyces cerevisiae PHO5 gene. Histone eviction from the PHO5 promoter during activation occurs with normal kinetics even in the absence of the PHO5 TATA box, showing that transcription of the gene itself is not required for promoter remodeling. Nevertheless, we find that mutations that impair transcript elongation by RNAPII affect the kinetics of histone eviction from the PHO5 promoter. Most dramatically, inactivation of RNAPII itself abolishes eviction completely. Under repressing conditions, an approximately 2.4-kb noncoding exosome-degraded transcript is detected that originates near the PHO5 termination site and is transcribed in the antisense direction. Abrogation of this transcript delays chromatin remodeling and subsequent RNAPII recruitment to PHO5 upon activation. We propose that noncoding transcription through positioned nucleosomes can enhance chromatin plasticity so that chromatin remodeling and activation of traversed genes occur in a timely manner.
RNA聚合酶II(RNAPII)进行的非编码转录,即基因间转录,在真核生物中极为普遍,但这种转录的作用仍知之甚少。在此我们表明,非编码转录在酿酒酵母PHO5基因的激活而非抑制中发挥作用。即使在没有PHO5 TATA框的情况下,激活过程中PHO5启动子上的组蛋白去除仍以正常动力学发生,这表明基因本身的转录对于启动子重塑并非必需。然而,我们发现损害RNAPII转录延伸的突变会影响PHO5启动子上组蛋白去除的动力学。最显著的是,RNAPII本身的失活会完全消除组蛋白去除。在抑制条件下,检测到一个约2.4 kb的非编码外切体降解转录本,其起源于PHO5终止位点附近,并以反义方向转录。该转录本的缺失会延迟激活时的染色质重塑以及随后RNAPII募集到PHO5。我们提出,通过定位核小体的非编码转录可以增强染色质可塑性,从而使染色质重塑和被转录基因的激活及时发生。