Effect of UTP sugar and base modifications on vaccinia virus early gene transcription.
Effect of UTP sugar and base modifications on vaccinia virus early gene transcription.
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UTP糖和碱基修饰对痘苗病毒早期基因转录的影响。
DOI：
10.1016/j.virol.2006.01.004
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Niles,EdwardG
中科院分区：
文献类型：
--
作者：
Mohamed,MohamedRagaa;Piacente,SarahC;Dickerman,Benjamin;Niles,EdwardG
先前的努力表明，RNA寡核苷酸含有转录终止信号UUUUUNU刺激提前终止牛痘病毒早期基因转录，在体外。这一观察结果表明，病毒转录终止可能是开发抗痘病毒药物的一个有吸引力的靶点。由于短RNA分子容易受到核酸酶消化的影响，因此它们的使用需要稳定化修饰。为了评估U 5 NU信号的核糖和尿嘧啶修饰对早期基因转录终止的影响，在体外牛痘病毒早期基因转录终止系统中检测了尿嘧啶碱基、核糖2′位和磷酸二酯键修饰的UTP衍生物。将4-S-U、5-methyl-U、2-S-U、pseudo U和2′-F-dU掺入到新生转录物中抑制转录终止。6-aza-U、2′-amino-U、2′-azido-U和2′-O-methyl-U抑制转录延长，导致短转录本的积累。大部分短的转录本保留在三元复合体中，并可被追踪成全长转录本。最初，测试终止信号中所有尿苷的衍生物。终止信号的部分修饰也降低了终止活性。在U9终止信号的前三个尿苷上引入2′-O甲基核糖降低了含有U9的寡核苷酸刺激体外转录终止的能力，进一步的修饰消除了这种活性。因此，病毒早期基因转录终止证明了对不能耐受碱基或糖修饰的U 5 NU信号的严格要求。此外，VTF显示通过模板中的T9序列增强转录延伸。这些结果表明，VTF除了在mRNA帽形成、早期基因转录终止和中间基因转录起始中起作用外，还可能在早期基因转录延伸中起微妙的作用。
Prior efforts demonstrated that RNA oligonucleotides containing the transcription termination signal UUUUUNU stimulate premature termination of vaccinia virus early gene transcription, in vitro. This observation suggests that viral transcription termination may be an attractive target for the development of anti-poxvirus agents. Since short RNA molecules are readily susceptible to nuclease digestion, their use would require stabilizing modifications. In order to evaluate the effect of both ribose and uracil modifications of the U5NU signal on early gene transcription termination, UTP derivatives harboring modifications to the uracil base, the 2′ position of the ribose sugar and the phosphodiester bond were examined in an in vitro vaccinia virus early gene transcription termination system. Incorporation of 4-S-U, 5-methyl-U, 2-S-U, pseudo U and 2′-F-dU into the nascent transcript inhibited transcription termination. 6-aza-U, 2′-amino-U, 2′-azido-U and 2′-O methyl-U inhibited transcription elongation resulting in the accumulation of short transcripts. The majority of the short transcripts remained in the ternary complex and could be chased into full-length transcripts. Initially, derivatives of all uridines in the termination signal were tested. Partial modification of the termination signal reduced termination activity, as well. Introduction of 2′-O methyl ribose to the first three uridines of the U9 termination signal reduced the ability of U9 containing oligonucleotides to stimulate in vitro transcription termination, in trans. Further modifications eliminated this activity. Thus, viral early gene transcription termination demonstrates a rigorous requirement for a U5NU signal that is unable to tolerate modification to the base or sugar. Additionally, VTF was shown to enhance transcription elongation through the T9 sequence in the template. These results suggest that VTF may play a subtle role in early gene transcription elongation in addition to its known function in mRNA cap formation, early gene transcription termination and intermediate gene transcription initiation.
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DOI：
10.1006/viro.1996.0177
发表时间：
1996
期刊：
Virology.
影响因子：
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作者：
Condit,RC;Lewis,JI;Quinn,M;Christen,LM;Niles,EG
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Niles,EG
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DOI：
10.1073/pnas.57.2.314
发表时间：
1967
影响因子：
11.1
作者：
BY J. R. Kates;B. R. TAIcAusLAN
通讯作者：
B. R. TAIcAusLAN
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影响因子：
4.8
作者：
M. Mohamed;E. G. Niles
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E. G. Niles
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DOI：
--
发表时间：
1992
影响因子：
11.1
作者：
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通讯作者：
S. Shuman
痘苗病毒核苷三磷酸磷酸水解酶 I 与线性寡核苷酸的相互作用。
DOI：
10.1021/bi9903749
发表时间：
1999
期刊：
Biochemistry.
影响因子：
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作者：
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通讯作者：
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