DNA轉(zhuǎn)錄為RNA包括三個階段:起始,,延伸,,終止,其中轉(zhuǎn)錄終止過程又可分為兩步,,RNA轉(zhuǎn)錄本的釋放和DNA模板的釋放,。II型RNA聚合酶負責(zé)真核生物體內(nèi)編碼蛋白質(zhì)的基因的轉(zhuǎn)錄,其轉(zhuǎn)錄終止也是蛋白質(zhì)表達過程中的重要一步,。
以往研究表明,,聚合酶II終止轉(zhuǎn)錄過程必需RNA 3'端的多聚腺苷酸信號〔poly(A)〕, RNA前體的轉(zhuǎn)錄后剪切加工也和轉(zhuǎn)錄終止過程相關(guān),。另外,在β球蛋白基因的的3'側(cè)還發(fā)現(xiàn)了專一性的轉(zhuǎn)錄終止信號,。這說明聚合酶II轉(zhuǎn)錄終止過程涉及多種因素,,但目前還不清楚這些因素間如何相互作用終止轉(zhuǎn)錄過程。
在最新研究中,,來自英國的科學(xué)家用一種簡單的核抽提液及自然的或人工合成的核酸模板監(jiān)測到了哺乳動物RNA聚合酶的轉(zhuǎn)錄終止過程,。結(jié)果表明,首先poly(A)位點的識別使轉(zhuǎn)錄終止成為可能,,此后poly(A)位點被剪切,,暴露出不受保護的RNA末端,,導(dǎo)致結(jié)合在聚合酶II上的RNA轉(zhuǎn)錄本在核酸外切酶的作用下降解,該降解過程促使DNA模板和聚合酶II的脫離,。而具有專一性終止信號的基因,,如β球蛋白基因,則是在終止信號轉(zhuǎn)錄后被剪切,,從而啟動了RNA轉(zhuǎn)錄本的降解,,致使DNA模板和聚合酶II脫離。而此終止過程也必須以poly(A)位點的識別為前提,。
該研究成果作為封面論文發(fā)表于2008年3月14日的《分子細胞》(Molecular Cell)上,。(科學(xué)網(wǎng) 穆宏平/編譯)
生物谷推薦原始出處:
(Molecular Cell),Vol 29, 600-610, 14 March 2008,,Steven West, Nicholas J. Proudfoot, and Michael J. Dye
Molecular Dissection of Mammalian RNA Polymerase II Transcriptional Termination
Steven West,1 Nicholas J. Proudfoot,1, and Michael J. Dye1,
1 Sir William Dunn School of Pathology, South Parks Road, Oxford OX1 3RE, UK
Summary
Transcriptional termination of mammalian RNA polymerase II (Pol II) is an essential but little-understood step in protein-coding gene expression. Mechanistically, termination by all DNA-dependent RNA polymerases can be reduced to two steps, namely release of the RNA transcript and release of the DNA template. Using a simple nuclear fractionation procedure, we have monitored transcript and template release in the context of both natural and artificial Pol II terminator sequences. We describe the timing and relationship between these events and in so doing establish the roles of the poly(A) signal, cotranscriptional RNA cleavage events, and 5′-3′ exonucleolytic RNA degradation in the mammalian Pol II termination process.