Uncovering protein conformational dynamics within two-component viral biomolecular condensates.

IF 4.5 3区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Protein Science Pub Date : 2025-07-01 DOI:10.1002/pro.70181
Alice Colyer, Julia Acker, Alexander Borodavka, Antonio N Calabrese
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引用次数: 0

Abstract

Biomolecular condensates selectively compartmentalize and organize biomolecules within the crowded cellular milieu and are instrumental in some disease mechanisms. Upon infection, many RNA viruses form biomolecular condensates that are often referred to as viral factories. The assembly mechanism of these viral factories remains poorly defined but involves transient, non-stoichiometric protein/RNA interactions, making their structural characterization challenging. Here, we sought to investigate the structural dynamics and intermolecular interactions of the key proteins responsible for condensate formation upon rotavirus infection, namely NSP2 (an RNA chaperone) and NSP5 (an intrinsically disordered protein [IDP]), using a combination of hydrogen-deuterium exchange mass spectrometry (HDX-MS), native MS, and biophysical tools. Our data reveal key structural features of intrinsically disordered NSP5 that are vital for condensate assembly and highlight inter/intra-protein interactions involved in condensate assembly. Moreover, we demonstrate that within a condensate there are altered conformational dynamics within the C-terminal region of NSP2, which has previously been shown to play a role in regulating its RNA chaperoning activity, and in the disordered regions of NSP5. We propose that altered conformational dynamics in NSP2 and NSP5 are critical for regulation of RNA annealing within a biomolecular condensate and for condensate assembly/client recruitment, respectively. Combined, our data demonstrate that the unique environment within a biomolecular condensate can tune functionally important protein conformational dynamics, which may play a crucial role in the replication of rotaviruses.

揭示双组分病毒生物分子凝聚物中的蛋白质构象动力学。
生物分子凝聚物在拥挤的细胞环境中选择性地分隔和组织生物分子,并在某些疾病机制中发挥重要作用。在感染后,许多RNA病毒形成生物分子凝聚体,通常被称为病毒工厂。这些病毒工厂的组装机制仍然不明确,但涉及瞬时的,非化学计量的蛋白质/RNA相互作用,使其结构表征具有挑战性。在这里,我们试图利用氢-氘交换质谱(HDX-MS)、天然质谱和生物物理工具的组合,研究轮状病毒感染时负责凝析物形成的关键蛋白,即NSP2(一种RNA伴侣)和NSP5(一种内在无序蛋白[IDP])的结构动力学和分子间相互作用。我们的数据揭示了内在无序的NSP5的关键结构特征,这些特征对凝析物的组装至关重要,并突出了凝析物组装中涉及的蛋白质间/蛋白质内相互作用。此外,我们还证明了在NSP2的c端区域和NSP5的无序区域内,凝聚物中存在改变的构象动力学,这在之前被证明在调节其RNA伴随活性中起作用。我们提出NSP2和NSP5构象动力学的改变分别对生物分子凝聚物中的RNA退火和凝聚物组装/客户端招募的调节至关重要。综上所述,我们的数据表明,生物分子凝聚体内的独特环境可以调节功能重要的蛋白质构象动力学,这可能在轮状病毒的复制中发挥关键作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Protein Science
Protein Science 生物-生化与分子生物学
CiteScore
12.40
自引率
1.20%
发文量
246
审稿时长
1 months
期刊介绍: Protein Science, the flagship journal of The Protein Society, is a publication that focuses on advancing fundamental knowledge in the field of protein molecules. The journal welcomes original reports and review articles that contribute to our understanding of protein function, structure, folding, design, and evolution. Additionally, Protein Science encourages papers that explore the applications of protein science in various areas such as therapeutics, protein-based biomaterials, bionanotechnology, synthetic biology, and bioelectronics. The journal accepts manuscript submissions in any suitable format for review, with the requirement of converting the manuscript to journal-style format only upon acceptance for publication. Protein Science is indexed and abstracted in numerous databases, including the Agricultural & Environmental Science Database (ProQuest), Biological Science Database (ProQuest), CAS: Chemical Abstracts Service (ACS), Embase (Elsevier), Health & Medical Collection (ProQuest), Health Research Premium Collection (ProQuest), Materials Science & Engineering Database (ProQuest), MEDLINE/PubMed (NLM), Natural Science Collection (ProQuest), and SciTech Premium Collection (ProQuest).
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