REDUNDANT ELEMENTS IN THE ADENOVIRUS TYPE-5 INVERTED TERMINAL REPEAT PROMOTE BIDIRECTIONAL TRANSCRIPTION INVITRO AND ARE IMPORTANT FOR VIRUS GROWTH-INVIVO
REDUNDANT ELEMENTS IN THE ADENOVIRUS TYPE-5 INVERTED TERMINAL REPEAT PROMOTE BIDIRECTIONAL TRANSCRIPTION INVITRO AND ARE IMPORTANT FOR VIRUS GROWTH-INVIVO
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DOI：
10.1016/0042-6822(91)90843-z
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发表时间：
1991-09-01
期刊：
影响因子：
3.7
通讯作者：
HEARING, P
中科院分区：
文献类型：
--
作者：
HATFIELD, L;HEARING, P
腺病毒末端反向重复序列（ITR）含有许多参与病毒DNA复制起始的顺式作用元件，以及多个与细胞转录因子SP1和ATF结合的基序。在这项研究中，我们利用Hela细胞的转录提取物来证明腺病毒5型ITR在体外促进双向转录。引物延伸分析表明，ITR指导转录起始位点内的末端重复序列和固定距离外的ITR。ITR也强烈刺激转录的早期区域1A（E1 A）的起始位点时，它位于紧接上游的E1 A TATA区。缺失和点突变分析表明，两个不同的顺式作用元件参与ITR依赖的体外转录活性。细胞转录因子SP1和ATF先前被证明与这两个区域结合。病毒体内突变分析表明，NFIII/OCT-1结合位点和保守的ATF基序对有效的病毒生长是重要的。ITR侧翼区的调控元件被发现在功能上取代这些位点。
The adenovirus inverted terminal repeat (ITR) contains a number ofcis-acting elements that are involved in the initiation of viral DNA replication, as well as multiple binding motifs for the cellular transcription factors SP1 and ATF. In this study, we utilized a Hela cell transcription extract to demonstrate that the adenovirus type 5 ITR promotes bidirectional transcriptionin vitro. Primer extension analyses demonstrated that the ITR directed transcription at initiation sites both within the terminal repeat and at fixed distances outside of the ITR. The ITR also strongly stimulated transcription at the early region 1A (E1A) initiation site when it was situated immediately upstream of the E1A TATA ☐ region. Deletion and point mutational analyses demonstrated that two distinctcis-acting elements were involved in these ITR-dependent transcriptional activitiesin vitro. Cellular transcription factors SP1 and ATF were previously shown to bind to these two regions. Analysis of viral mutantsin vivodemonstrated that the NFIII/OCT-1 binding site and a conserved ATF motif were important for efficient viral growth. Regulatory elements in the ITR flanking region were found to functionally substitute for these sites.