RNA聚合酶识别启动子.ppt

大肠杆菌RNA聚合酶: a 亚基 在核心酶中两个a亚基是相同的； a亚基是由rpoA 基因编码； 对于RNA聚合酶核心蛋白的组装是必需的； 在启动子识别上可能起着重要的作用； 1. β亚基由rpoB 基因编码； 2. RNA聚合酶的催化中心； Rifampicin (利福平)：与β亚基结合可以抑制转录的起始，rpoB 基因突变导致对利福平的抗性； Streptolydigins(利迪链菌素)：抗性突变也定位于 rpoB 基因，它抑制转录延伸，但不抑制起始； 3. β亚基可能含有两个结构域负责转录的起始和延伸； 1. b 亚基由rpoC 基因编码； 2. 与两个Zn2+ 离子结合参与RNA聚合酶的催化功能； Heparin (肝素)：在体外与 β’ 亚基结合并抑制转录； Heparin 与DNA竞争结合RNA聚合酶； 3. β’ 亚基可能是负责与DNA模板的结合； 1. 在启动子的识别上σ 因子至关重要，对于非特异性DNA位点核心酶的亲和力会降低（104），而对于相应的启动子亲和力会增加； 2. 当起始后(RNA chain is 8-9 nt) ，σ 因子会从RNA聚合酶中释放出来； 3. 在细胞中σ 因子的需要量比RNA聚合酶其他亚基的需要量少； 转录起始点 转录起始位点没有广泛的序列同源性，但第一个碱基为腺嘌呤，而两侧是嘧啶碱基。这个区域被称为起始子（initiator， Inr），序列可表示为PyPyANT/APyPy。Inr元件位于-2～+4。仅由Inr元件组成的启动子是具有可被RNA聚合酶II识别的最简单启动子形式 转录起始位点与TATA box一起组成核心启动子； 上游调控元件具有促进转录作用 TFIIIB: (1) binds 50 bp upstream from the A box, but has no sequence specificity and the binding position is determined by the DNA bound TFIIIC. (2) consists of three subunits, one of which is TBP, the general initiation factor; the second is called BRF (TFIIB-related factor); and the third is called B”. TFIIIC: A and B boxes binding and a assembly factor to position TFIIIB TFIIIB: DNA binding and RNA Pol III recruiting 5.7-3 5S rRNA genes Tandemly arranged in a gene cluster. (In human, there is a single cluster of around 2000 genes.) Transcription control regions (promoters) are organized similar to those of tRNA, except that C box is in place of B box. C box: +81-99 bp; A box: +50-65 3. Transcription factors: (1) The C box acts as the binding site for TFIIIA. (2) TFIIIA acts as an assembly factor which allows TFIIIC to interact with the 5S rRNA promoter. (3) The A box may also stabilize TFIIIC binding. (4) TFIIIC is then bound to DNA site near +1. (5) TFIIIB and TFIIIC interact to recruit RNA Pol III to initiate transcription. TFIIIA TFIIIC TFIIIB Pol III 5.7-4 Alternative RNA Pol III promoters Many RNA Pol III genes also rely on upstream sequences for regulation of their transcription e.g. U6 snRNA and Epstein-Barr virus Use only re