Capping of vesicular stomatitis virus pre-mRNA is required for accurate selection of transcription stop-start sites and virus propagation.
Capping of vesicular stomatitis virus pre-mRNA is required for accurate selection of transcription stop-start sites and virus propagation.
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DOI：
10.1093/nar/gku901
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发表时间：
2014-10-29
影响因子：
14.9
通讯作者：
Ogino T
中科院分区：
文献类型：
--
作者：
Ogino T
水泡性口炎病毒的多功能RNA依赖性RNA聚合酶L蛋白通过共价酶-pRNA中间体的形成催化非常规的前mRNA加帽，这需要多聚核糖核苷酸转移酶结构域中的组氨酸-精氨酸(HR)基序。在这里，使用体外重建转录系统分析了 HR 基序中帽子缺陷突变对转录的影响。野生型 L 蛋白通过停止-启动转录机制从基因组 3' 端合成前导 RNA，然后从内部基因合成 5' 加帽和 3'-多腺苷酸化 mRNA。帽缺陷突变体有效地产生了前导RNA，但使用第一个基因内的隐性终止和起始信号显示出异常的停止-起始转录，导致从正确的mRNA起始位点顺序生成带有5'-ATP的~40-核苷酸转录本，然后是从非典型起始位点用非规范GTP起始的28-核苷酸转录本和长3'-聚腺苷酸化转录本。第一个基因内频繁的转录终止和重新启动显着减弱了下游 mRNA 的产生。与这些突变体无法进行体外 mRNA 合成和加帽一致，这些突变对于培养细胞中的病毒复制是致命的。这些发现表明，病毒 mRNA 加帽是准确停止-启动转录以及 mRNA 稳定性和翻译所必需的，因此也是病毒在宿主细胞中复制所必需的。
The multifunctional RNA-dependent RNA polymerase L protein of vesicular stomatitis virus catalyzes unconventional pre-mRNA capping via the covalent enzyme-pRNA intermediate formation, which requires the histidine–arginine (HR) motif in the polyribonucleotidyltransferase domain. Here, the effects of cap-defective mutations in the HR motif on transcription were analyzed using an in vitro reconstituted transcription system. The wild-type L protein synthesized the leader RNA from the 3′-end of the genome followed by 5′-capped and 3′-polyadenylated mRNAs from internal genes by a stop–start transcription mechanism. Cap-defective mutants efficiently produced the leader RNA, but displayed aberrant stop–start transcription using cryptic termination and initiation signals within the first gene, resulting in sequential generation of ∼40-nucleotide transcripts with 5′-ATP from a correct mRNA-start site followed by a 28-nucleotide transcript and long 3′-polyadenylated transcript initiated with non-canonical GTP from atypical start sites. Frequent transcription termination and re-initiation within the first gene significantly attenuated the production of downstream mRNAs. Consistent with the inability of these mutants in in vitro mRNA synthesis and capping, these mutations were lethal to virus replication in cultured cells. These findings indicate that viral mRNA capping is required for accurate stop–start transcription as well as mRNA stability and translation and, therefore, for virus replication in host cells.
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影响因子：
5.4
作者：
Barr, JN;Whelan, SPJ;Wertz, GW
通讯作者：
Wertz, GW
DOI：
10.1016/0006-291x(85)90568-6
发表时间：
1985-01-01
影响因子：
3.1
作者：
DE, BP;BANERJEE, AK
通讯作者：
BANERJEE, AK
影响因子：
5.4
作者：
Li, Jianrong;Rahmeh, Amal;Whelan, Sean P. J.
通讯作者：
Whelan, Sean P. J.
DOI：
10.1073/pnas.92.10.4477
发表时间：
1995-05-09
影响因子：
11.1
作者：
LAWSON, ND;STILLMAN, EA;ROSE, JK
通讯作者：
ROSE, JK
DOI：
10.1073/pnas.70.12.3566
发表时间：
1973-01-01
影响因子：
11.1
作者：
BANERJEE, AK;RHODES, DP
通讯作者：
RHODES, DP