Structural insight and flexible features of NS5 proteins from all four serotypes of Dengue virus in solution.

IF 2.1 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Biotechnology Letters Pub Date : 2015-11-01 Epub Date: 2015-10-31 DOI:10.1107/S1399004715017721

Wuan Geok Saw, Giancarlo Tria, Ardina Grüber, Malathy Sony Subramanian Manimekalai, Yongqian Zhao, Arun Chandramohan, Ganesh Srinivasan Anand, Tsutomu Matsui, Thomas M Weiss, Subhash G Vasudevan, Gerhard Grüber

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Abstract

Infection by the four serotypes of Dengue virus (DENV-1 to DENV-4) causes an important arthropod-borne viral disease in humans. The multifunctional DENV nonstructural protein 5 (NS5) is essential for capping and replication of the viral RNA and harbours a methyltransferase (MTase) domain and an RNA-dependent RNA polymerase (RdRp) domain. In this study, insights into the overall structure and flexibility of the entire NS5 of all four Dengue virus serotypes in solution are presented for the first time. The solution models derived revealed an arrangement of the full-length NS5 (NS5FL) proteins with the MTase domain positioned at the top of the RdRP domain. The DENV-1 to DENV-4 NS5 forms are elongated and flexible in solution, with DENV-4 NS5 being more compact relative to NS5 from DENV-1, DENV-2 and DENV-3. Solution studies of the individual MTase and RdRp domains show the compactness of the RdRp domain as well as the contribution of the MTase domain and the ten-residue linker region to the flexibility of the entire NS5. Swapping the ten-residue linker between DENV-4 NS5FL and DENV-3 NS5FL demonstrated its importance in MTase-RdRp communication and in concerted interaction with viral and host proteins, as probed by amide hydrogen/deuterium mass spectrometry. Conformational alterations owing to RNA binding are presented.

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四种血清型登革热病毒 NS5 蛋白在溶液中的结构洞察力和柔性特征。

登革热病毒（DENV-1 至 DENV-4）的四个血清型会导致人类感染一种重要的节肢动物传播病毒性疾病。多功能 DENV 非结构蛋白 5（NS5）对于病毒 RNA 的封顶和复制至关重要，它含有一个甲基转移酶（MTase）结构域和一个 RNA 依赖性 RNA 聚合酶（RdRp）结构域。本研究首次揭示了四种登革热病毒血清型的整个 NS5 在溶液中的整体结构和灵活性。所得到的溶液模型揭示了全长 NS5（NS5FL）蛋白的排列，MTase 结构域位于 RdRP 结构域的顶部。从DENV-1到DENV-4的NS5在溶液中都是伸长而灵活的，其中DENV-4的NS5相对于DENV-1、DENV-2和DENV-3的NS5更为紧凑。对单个MTase和RdRp结构域的溶液研究显示了RdRp结构域的紧凑性以及MTase结构域和十个残基连接区对整个NS5灵活性的贡献。将DENV-4 NS5FL和DENV-3 NS5FL之间的十个残基连接区互换后，通过酰胺氢/氘质谱分析，证明了其在MTase-RdRp沟通以及与病毒和宿主蛋白协同作用方面的重要性。报告介绍了 RNA 结合导致的构象变化。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.