Analysis of flavivirus NS5 methyltransferase cap binding.
Analysis of flavivirus NS5 methyltransferase cap binding.
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DOI：
10.1016/j.jmb.2008.11.058
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发表时间：
2009-02-06
影响因子：
5.6
通讯作者：
Peersen, Olve B.
中科院分区：
文献类型：
--
作者：
Geiss, Brian J.;Thompson, Aaron A.;Andrews, Andrew J.;Sons, Robert L.;Gari, Hamid H.;Keenan, Susan M.;Peersen, Olve B.
黄病毒NS 5 N-末端RNA甲基转移酶（MTase）负责甲基化病毒RNA帽结构。为了增加我们对病毒RNA帽结合机制的理解，我们对登革热（DEN）和黄热病（YF）病毒MTase酶的鸟苷帽结合口袋进行了详细的结构和生化表征。我们解析了DEN 2 MTase的改进的2.1 nm分辨率晶体结构和载脂蛋白形式的YF病毒MTase结构域的新的1.5 nm分辨率晶体结构以及与GTP和RNA帽类似物复合的新的1.45 nm结构。我们的结构澄清了以前报道的DEN MTase结构，提出了新的蛋白质帽相互作用，并提供了鸟嘌呤特异性的详细视图。此外，DEN和YF蛋白质的结构基本上是相同的，表明黄病毒MT酶之间存在很大程度的结构保守性。进行GTP类似物竞争测定和诱变分析以分析帽结合的生物化学特征，确定主要的相互作用点是（i）鸟嘌呤环通过与Phe 24的π-π堆积，N1氢通过水桥与Leu 19骨架羰基相互作用，以及C2胺与Leu 16和Leu 19骨架羰基相互作用，（ii）核糖2′-羟基与Lys 13和Asn 17的相互作用，以及（iii）α-磷酸与Lys 28和Ser 215的相互作用。基于我们的突变和类似物研究，鸟嘌呤环和α-磷酸相互作用提供了帽结合的大部分能量，而鸟嘌呤N1和Leu 19羰基之间的水桥和C2胺和Leu 16/Leu 19羰基之间的氢键的组合提供了特异性鸟嘌呤识别。黄病毒MTase蛋白如何结合RNA帽结构的详细模型。
The flavivirus NS5 N-terminal RNA methyltransferase (MTase) enzyme is responsible for methylating the viral RNA cap structure. To increase our understanding of the mechanism of viral RNA cap binding we performed a detailed structural and biochemical characterization of guanosine cap binding pocket of the dengue (DEN) and yellow fever (YF) virus MTase enzymes. We solved an improved 2.1 Å resolution crystal structure of DEN2 MTase and new 1.5 Å resolution crystal structures of the YF virus MTase domain in apo form and new 1.45 Å structure in complex with GTP and RNA cap analog. Our structures clarify the previously reported DEN MTase structure, suggest novel protein–cap interactions, and provide a detailed view of guanine specificity. Furthermore, the structures of the DEN and YF proteins are essentially identical, indicating a large degree of structural conservation amongst the flavivirus MTases. GTP analog competition assays and mutagenesis analysis, performed to analyze the biochemical characteristics of cap binding, determined that the major interaction points are (i) guanine ring via π–π stacking with Phe24, N1 hydrogen interaction with the Leu19 backbone carbonyl via a water bridge, and C2 amine interaction with Leu16 and Leu19 backbone carbonyls, (ii) ribose 2′ hydroxyl interaction with Lys13 and Asn17, and (iii) α-phosphate interactions with Lys28 and Ser215. Based on our mutational and analog studies, the guanine ring and α-phosphate interactions provide most of the energy for cap binding, while the combination of the water bridge between the guanine N1 and Leu19 carbonyl and the hydrogen bonds between the C2 amine and Leu16/Leu19 carbonyl groups provide for specific guanine recognition. A detailed model of how the flavivirus MTase protein binds RNA cap structures is presented.
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影响因子：
3.8
作者：
Assenberg, Rene;Ren, Jingshan;Grimes, Jonathan M.
通讯作者：
Grimes, Jonathan M.
影响因子：
3.8
作者：
Kuo, MD;Chin, C;Lin, JH
通讯作者：
Lin, JH
影响因子：
11.4
作者：
Mazza, C;Segref, A;Cusack, S
通讯作者：
Cusack, S
影响因子：
5.8
作者：
Theobald, Douglas L.;Wuttke, Deborah S.
通讯作者：
Wuttke, Deborah S.
DOI：
10.1107/s0907444998003254
发表时间：
1998-09-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
Brunger, AT;Adams, PD;Warren, GL
通讯作者：
Warren, GL