The N-terminal domain of the arenavirus L protein is an RNA endonuclease essential in mRNA transcription.
The N-terminal domain of the arenavirus L protein is an RNA endonuclease essential in mRNA transcription.
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DOI：
10.1371/journal.ppat.1001038
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发表时间：
2010-09-16
期刊：
影响因子：
6.7
通讯作者：
Canard B
中科院分区：
文献类型：
--
作者：
Morin B;Coutard B;Lelke M;Ferron F;Kerber R;Jamal S;Frangeul A;Baronti C;Charrel R;de Lamballerie X;Vonrhein C;Lescar J;Bricogne G;Günther S;Canard B
沙粒病毒利用短帽引物合成病毒mrna，这可能是通过“帽抢夺”机制从细胞转录本中获得的。在这里，我们报告了原型沙粒病毒：淋巴细胞性脉络丛脑膜炎病毒L蛋白n端196个残基（NL1）的晶体结构和功能表征。NL1结构域能够结合和切割RNA。NL1的2.13 Å分辨率晶体结构揭示了类似于流感病毒PA蛋白n末端的II型内切酶α/β结构。两种结构的叠加、诱变和反向遗传学研究揭示了PD…（D/E）XK II型内切酶特征序列的关键活性位点残基的独特空间排列。我们发现这个核酸内切酶结构域在沙粒病毒科、布尼亚病毒科和正粘病毒科中都是保守的和活跃的，并提出沙粒病毒NL1结构域是沙粒病毒的夺帽核酸内切酶。沙粒病毒家族包括几种危及生命的人类病原体，可引起脑膜炎或出血热。这些RNA病毒利用一种目前没有结构数据的RNA合成机制复制和转录它们的基因组。他们使用短帽引物合成病毒mrna，这些引物可能是通过一种“帽抢夺”机制从细胞转录本中获得的，这种机制被认为涉及携带RNA依赖性RNA聚合酶特征序列的大L蛋白。在这里，我们报道了从原型沙粒病毒：淋巴细胞性脉络丛脑膜炎病毒中分离的L蛋白（NL1） n端结构域的晶体结构和功能表征。NL1结构域能够结合和切割RNA。NL1的2.13 Å分辨率晶体结构揭示了类似于流感病毒PA蛋白n末端的II型内切酶α/β结构。两种结构的叠加和诱变研究揭示了PD…（D/E）XK II型内切酶特征序列相关的关键活性位点残基的独特空间排列。反向遗传研究表明，活性位点残基的突变选择性地终止转录，而不是复制。我们发现该内切酶结构域在沙粒病毒科、布尼亚病毒科和正粘病毒科中是保守的和活跃的，并提出沙粒病毒NL1结构域是沙粒病毒的夺帽内切酶。
Arenaviridae synthesize viral mRNAs using short capped primers presumably acquired from cellular transcripts by a ‘cap-snatching’ mechanism. Here, we report the crystal structure and functional characterization of the N-terminal 196 residues (NL1) of the L protein from the prototypic arenavirus: lymphocytic choriomeningitis virus. The NL1 domain is able to bind and cleave RNA. The 2.13 Å resolution crystal structure of NL1 reveals a type II endonuclease α/β architecture similar to the N-terminal end of the influenza virus PA protein. Superimposition of both structures, mutagenesis and reverse genetics studies reveal a unique spatial arrangement of key active site residues related to the PD…(D/E)XK type II endonuclease signature sequence. We show that this endonuclease domain is conserved and active across the virus families Arenaviridae, Bunyaviridae and Orthomyxoviridae and propose that the arenavirus NL1 domain is the Arenaviridae cap-snatching endonuclease. The Arenaviridae virus family includes several life-threatening human pathogens that cause meningitis or hemorrhagic fever. These RNA viruses replicate and transcribe their genome using an RNA synthesis machinery for which no structural data currently exist. They synthesize viral mRNAs using short capped primers presumably acquired from cellular transcripts by a ‘cap-snatching’ mechanism thought to involve the large L protein, which carries RNA-dependent RNA polymerase signature sequences. Here, we report the crystal structure and functional characterization of an isolated N-terminal domain of the L protein (NL1) from the prototypic arenavirus: lymphocytic choriomeningitis virus. The NL1 domain is able to bind and cleave RNA. The 2.13 Å resolution crystal structure of NL1 reveals a type II endonuclease α/β architecture similar to the N-terminal end of the influenza virus PA protein. Superimposition of both structures and mutagenesis studies reveal a unique spatial arrangement of key active site residues related to the PD…(D/E)XK type II endonuclease signature sequence. Reverse genetic studies show that mutation of active site residues selectively abolish transcription, not replication. We show that this endonuclease domain is conserved and active across the virus families: Arenaviridae, Bunyaviridae and Orthomyxoviridae and propose that the arenavirus NL1 domain is the Arenaviridae cap-snatching endonuclease.
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DOI：
10.1107/s0907444905036693
发表时间：
2006-01-01
影响因子：
2.2
作者：
Evans, P
通讯作者：
Evans, P
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
5.4
作者：
GARCIN, D;KOLAKOFSKY, D
通讯作者：
KOLAKOFSKY, D
影响因子：
3.6
作者：
BARTON, LL;BUDD, SC;YOSHINO, MT
通讯作者：
YOSHINO, MT
影响因子：
64.8
作者：
Dias, Alexandre;Bouvier, Denis;Ruigrok, Rob W. H.
通讯作者：
Ruigrok, Rob W. H.