Synthesis of novel products in vitro by an RNA-dependent RNA polymerase
Synthesis of novel products in vitro by an RNA-dependent RNA polymerase
复制标题
通过 RNA 依赖性 RNA 聚合酶体外合成新产品
作者：
C. Song;A. Simon
芜菁皱缩病毒感染芜菁的RNA依赖性RNA聚合酶在体外转录病毒相关卫星RNA sat-RNA C（356个碱基）的两条链。虽然正链和负链sat-RNA C都可以指导全长互补链产物的合成，但负链RNA的转录也产生两种非模板大小的产物，L-RNA和S-RNA（C. Song和A. E. Simon，Proc. Natl. Acad. Sci. USA 91：8792-8796，1994）。本文报道了L-RNA和S-RNA的合成是由模板3'末端的末端延伸引起的。L-RNA具有柄状结构，并且由与新合成的RNA共价连接的负链模板组成，所述新合成的RNA与其5' 190个碱基互补。S-RNA是由模板与其全长互补链共价连接而成。所有检测的负链模板均产生S-RNA。然而，L-RNA的合成受到负链模板的3'端或几个内部区域的缺失以及5'端附近或紧邻模板-产物连接上游的内部序列中的碱基改变的影响，这些碱基改变可能与3'端形成异源双链体。此外，破坏或恢复参与体内RNA重组的茎环的突变影响了体外产生的L-RNA的水平，这表明分子内形成柄状RNA和分子间RNA重组的机制涉及类似的特征。
RNA-dependent RNA polymerase from turnip crinkle virus-infected turnip transcribes both strands of a virus-associated satellite RNA, sat-RNA C (356 bases), in vitro. While both plus- and minus-strand sat-RNA C can direct the synthesis of full-length complementary-strand products, transcription of minus-strand RNA also generates two non-template-sized products, L-RNA and S-RNA (C. Song and A. E. Simon, Proc. Natl. Acad. Sci. USA 91:8792-8796, 1994). Here we report that synthesis of L-RNA and S-RNA results from terminal elongation of the 3' end of the template. L-RNA has a panhandle structure and is composed of minus-strand template covalently linked to newly synthesized RNA complementary to its 5' 190 bases. S-RNA is composed of template covalently linked to its full-length complementary strand. All minus-strand templates tested yielded S-RNA. However, synthesis of L-RNA was affected by deletion of the 3' end of the minus-strand template or several internal regions and base alterations near the 5' end or in an internal sequence immediately upstream from the template-product junction that could potentially form a heteroduplex with the 3' end. Furthermore, mutations that disrupted or restored a stem-loop involved in RNA recombination in vivo affected the level of L-RNA produced in vitro, suggesting that the mechanisms for intramolecular formation of panhandle RNAs and intermolecular RNA recombination involve similar features.
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DOI：
10.1073/pnas.85.23.8998
发表时间：
1988-12-01
影响因子：
11.1
作者：
FROHMAN, MA;DUSH, MK;MARTIN, GR
通讯作者：
MARTIN, GR
Q beta 复制酶合成 RNA 时的末端腺苷酸化。
DOI：
--
发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Bausch,JN;Kramer,FR;Miele,EA;Dobkin,C;Mills,DR
通讯作者：
Mills,DR
DOI：
10.1073/pnas.90.4.1498
发表时间：
1993-02-15
影响因子：
11.1
作者：
QUADT, R;KAO, CC;AHLQUIST, P
通讯作者：
AHLQUIST, P
影响因子：
14.9
作者：
MILLIGAN, JF;GROEBE, DR;UHLENBECK, OC
通讯作者：
UHLENBECK, OC
来自黑甲虫病毒感染的果蝇细胞的模板依赖性 RNA 聚合酶。
影响因子：
3.7
作者：
Saunders,K;Kaesberg,P
通讯作者：
Kaesberg,P