PABP-driven secondary condensed phase within RSV inclusion bodies activates viral mRNAs for ribosomal recruitment

Qiang Zhang; Hanzhe Ye; Cong Liu; Haiwu Zhou; Mingbin He; Xiaodong Liang; Yu Zhou; Kun Wang; Yali Qin; Zhifei Li; Mingzhou Chen

doi:10.1016/j.virs.2023.12.001

April 2024

. doi: 10.1016/j.virs.2023.12.001

Citation: Qiang Zhang, Hanzhe Ye, Cong Liu, Haiwu Zhou, Mingbin He, Xiaodong Liang, Yu Zhou, Kun Wang, Yali Qin, Zhifei Li, Mingzhou Chen. PABP-driven secondary condensed phase within RSV inclusion bodies activates viral mRNAs for ribosomal recruitment .VIROLOGICA SINICA, 2024, 39(2) : 235-250. http://dx.doi.org/10.1016/j.virs.2023.12.001

呼吸道合胞病毒包涵体内PABP驱动的次级凝聚相调控病毒mRNAs的活化

张强 ^a ,
叶瀚哲 ^a ,
刘聪 ^a ,
周海武 ^a ,
何铭斌 ^a ,
梁晓东 ^a ,
周宇 ^a ,
王坤 ^a ,
覃雅丽 ^a,, ,
李智菲 ^a,, ,
陈明周 ^a,b,c,,

a. 武汉大学病毒学国家重点实验室;
b. 武汉大学泰康生命医学中心;
c. 湖北江夏实验室

通讯作者： 覃雅丽, yqin@whu.edu.cn; 李智菲, zhifeili@whu.edu.cn; 陈明周, chenmz@whu.edu.cn
收稿日期： 2023-11-18
录用日期： 2023-12-03

摘要

呼吸道合胞病毒（Respiratory syncytial virus, RSV）是引起全世界婴幼儿急性下呼吸道感染（acute lower respiratory tract infection, ALRTI）最重要的病原体。RSV的包涵体（inclusion bodies, IBs）通过液-液相分离（liquid-liquid phase separation, LLPS）形成, 其内部结构——包涵体相关颗粒（IB-associated granules, IBAGs），瞬时浓缩新合成的病毒mRNAs及转录抗终止因子M2-1，但是IBAGs的形成机制以及如何调控病毒mRNAs翻译的分子机制仍不明确。本研究发现RSV IBs的内部结构实际上是由次级LLPS形成的、不含M2-1的病毒信使核糖核蛋白（messenger ribonucleoprotein, mRNP）凝聚物。机制上，RSV核蛋白（nucleoprotein, N）和M2-1与PABP相互作用将PABP募集至RSV IBs，促使PABP通过RNA识别基序（RNA-recognition motif, RRM）结合IBs内转录的病毒mRNAs并驱动次级相分离，即PABP是驱动次级相分离的支架蛋白。此外，PABP-eIF4G1相互作用调控病毒mRNP凝聚物的组成，将特异的翻译起始因子（eIF4G1、eIF4E、eIF4A、eIF4B 呼吸道合胞病毒（Respiratory syncytial virus, RSV）是引起全世界婴幼儿急性下呼吸道感染（acute lower respiratory tract infection, ALRTI）最重要的病原体。RSV的包涵体（inclusion bodies, IBs）通过液-液相分离（liquid-liquid phase separation, LLPS）形成, 其内部结构——包涵体相关颗粒（IB-associated granules, IBAGs），瞬时浓缩新合成的病毒mRNAs及转录抗终止因子M2-1，但是IBAGs的形成机制以及如何调控病毒mRNAs翻译的分子机制仍不明确。本研究发现RSV IBs的内部结构实际上是由次级LLPS形成的、不含M2-1的病毒信使核糖核蛋白（messenger ribonucleoprotein, mRNP）凝聚物。机制上，RSV核蛋白（nucleoprotein, N）和M2-1与PABP相互作用将PABP募集至RSV IBs，促使PABP通过RNA识别基序（RNA-recognition motif, RRM）结合IBs内转录的病毒mRNAs并驱动次级相分离，即PABP是驱动次级相分离的支架蛋白。此外，PABP-eIF4G1相互作用调控病毒mRNP凝聚物的组成，将特异的翻译起始因子（eIF4G1、eIF4E、eIF4A、eIF4B和eIF4H）募集至次级凝聚相中。最后，本研究发现病毒mRNP凝聚物的功能是对病毒mRNAs进行活化，促进病毒mRNAs对核糖体的募集，从而提高病毒mRNAs的翻译效率。我们的研究揭示了一种新的、由LLPS调控的病毒蛋白翻译机制，提供了一种新的靶向次级凝聚相的抗病毒策略。
- 呼吸道合胞病毒
- , 生物分子凝聚物
- , 包涵体
- , 液-液相分离
- , 翻译
- , mRNA活化

PABP-driven secondary condensed phase within RSV inclusion bodies activates viral mRNAs for ribosomal recruitment

Qiang Zhang ^a ,
Hanzhe Ye ^a ,
Cong Liu ^a ,
Haiwu Zhou ^a ,
Mingbin He ^a ,
Xiaodong Liang ^a ,
Yu Zhou ^a ,
Kun Wang ^a ,
Yali Qin ^a,, ,
Zhifei Li ^a,, ,
Mingzhou Chen ^a,b,c,,

a. State Key Laboratory of Virology and Modern Virology Research Center, College of Life Sciences, Wuhan University, Wuhan, 430072, China;
b. Taikang Center for Life and Medical Sciences, Wuhan University, Wuhan, 430072, China;
c. Hubei Jiangxia Laboratory, Wuhan, 430200, China

Corresponding author: Yali Qin, yqin@whu.edu.cn Zhifei Li, zhifeili@whu.edu.cn Mingzhou Chen, chenmz@whu.edu.cn
Received Date: 18 November 2023
Accepted Date: 03 December 2023

Abstract

Inclusion bodies (IBs) of respiratory syncytial virus (RSV) are formed by liquid-liquid phase separation (LLPS) and contain internal structures termed “IB-associated granules” (IBAGs), where anti-termination factor M2-1 and viral mRNAs are concentrated. However, the mechanism of IBAG formation and the physiological function of IBAGs are unclear. Here, we found that the internal structures of RSV IBs are actual M2-1-free viral messenger ribonucleoprotein (mRNP) condensates formed by secondary LLPS. Mechanistically, the RSV nucleoprotein (N) and M2-1 interact with and recruit PABP to IBs, promoting PABP to bind viral mRNAs transcribed in IBs by RNA-recognition motif and drive secondary phase separation. Furthermore, PABP-eIF4G1 interaction regulates viral mRNP condensate composition, thereby recruiting specific translation initiation factors (eIF4G1, eIF4E, eIF4A, eIF4B and eIF4H) into the secondary condensed phase to activate viral mRNAs for ribosomal recruitment. Our study proposes a novel LLPS-regulated translation mechanism during viral infection and a novel antiviral strategy via targeting on secondary condensed phase.
- Respiratory syncytial virus (RSV)
- , Inclusion bodies (IBs)
- , Biomolecular condensates
- , Liquid-liquid phase separation (LLPS)
- , Secondary condensed phase
- , mRNA activation

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