Laboratory of Transcription Biology, Molecular Genetics Division, CSIR-Indian Institute of Chemical Biology, 4 Raja, S.C. Mullick Road, Kolkata 32, India.
Laboratory of Biochemistry and Molecular Biology, The Rockefeller University, New York, NY 10065, USA.
Cell Rep. 2019 Jan 29;26(5):1303-1317.e7. doi: 10.1016/j.celrep.2019.01.012.
Despite substantial progress in our understanding of the players involved and the regulatory mechanisms controlling the initiation and elongation steps of transcription, little is known about the recruitment of elongation factors at promoter-proximal regions for the initiation-to-elongation transition. Here, we show evidence that human TFIID, which initiates pre-initiation complex (PIC) assembly, contributes to regulating the recruitment of super-elongation complex (SEC) components at the promoter-proximal region through interactions among selective TAF and SEC components. In vitro direct interactions, coupled with cell-based assays, identified an important poly-Ser domain within SEC components that are involved in their interaction with TFIID. DNA template-based recruitment assays, using purified components, further show a direct role for poly-Ser domain-dependent TFIID interaction in recruiting SEC components on target DNA. Consistently, ChIP and RNA analyses have shown the importance of this mechanism in TFIID-dependent SEC recruitment and target gene expression within mammalian cells.
尽管我们在理解参与的参与者和控制转录起始和延伸步骤的调节机制方面取得了重大进展,但对于起始到延伸转变过程中在启动子近端区域募集延伸因子的情况知之甚少。在这里,我们表明,起始前复合物 (PIC) 组装的人 TFIID 通过选择性 TAF 和 SEC 成分之间的相互作用,有助于调节启动子近端区域中超延伸复合物 (SEC) 成分的募集。体外直接相互作用结合基于细胞的测定,鉴定了 SEC 成分中涉及与 TFIID 相互作用的重要多丝氨酸结构域。使用纯化成分的基于 DNA 模板的募集测定进一步表明,多丝氨酸结构域依赖性 TFIID 相互作用在募集靶 DNA 上的 SEC 成分中的直接作用。一致地,ChIP 和 RNA 分析表明,在哺乳动物细胞中,该机制在 TFIID 依赖性 SEC 募集和靶基因表达中具有重要意义。