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人类巨细胞病毒的二次包膜是一个快速过程，利用内吞体作为主要的膜源。

Secondary Envelopment of Human Cytomegalovirus Is a Fast Process Utilizing the Endocytic Compartment as a Major Membrane Source.

机构信息

Central Facility for Electron Microscopy, Ulm University, 89081 Ulm, Germany.

Institute of Virology, Ulm University Medical Center, 89081 Ulm, Germany.

出版信息

Biomolecules. 2024 Sep 12;14(9):1149. doi: 10.3390/biom14091149.

DOI:10.3390/biom14091149
PMID:39334915
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11430300/
Abstract

Secondary envelopment of the human cytomegalovirus (HCMV) is a critical but not well-understood process that takes place at the cytoplasmic viral assembly complex (cVAC) where nucleocapsids acquire their envelope by budding into cellular membranes containing viral glycoproteins. Previous studies presented controversial results regarding the composition of the viral envelope, suggesting trans-Golgi and endosomal origins, as well as intersections with the exosomal and endocytic pathways. Here, we investigated the role of endocytic membranes for the secondary envelopment of HCMV by using wheat germ agglutinin (WGA) pulse labeling to label glycoproteins at the plasma membrane and to follow their trafficking during HCMV infection by light microscopy and transmission electron microscopy (TEM). WGA labeled different membrane compartments within the cVAC, including early endosomes, multivesicular bodies, trans-Golgi, and recycling endosomes. Furthermore, TEM analysis showed that almost 90% of capsids undergoing secondary envelopment and 50% of enveloped capsids were WGA-positive within 90 min. Our data reveal extensive remodeling of the endocytic compartment in the late stage of HCMV infection, where the endocytic compartment provides an optimized environment for virion morphogenesis and serves as the primary membrane source for secondary envelopment. Furthermore, we show that secondary envelopment is a rapid process in which endocytosed membranes are transported from the plasma membrane to the cVAC within minutes to be utilized by capsids for envelopment.

摘要

人巨细胞病毒 (HCMV) 的二次包膜是一个关键但尚未被充分理解的过程，该过程发生在细胞质病毒组装复合物 (cVAC) 中，核衣壳通过出芽进入含有病毒糖蛋白的细胞膜获得包膜。先前的研究对病毒包膜的组成提出了争议的结果，表明其具有反式高尔基体和内体起源，以及与外泌体和内吞途径的交叉。在这里，我们通过使用麦胚凝集素 (WGA) 脉冲标记来标记质膜上的糖蛋白，并通过荧光显微镜和透射电子显微镜 (TEM) 观察 HCMV 感染过程中糖蛋白的运输，来研究内吞膜在 HCMV 二次包膜中的作用。WGA 标记了 cVAC 内的不同膜区室，包括早期内体、多泡体、反式高尔基体和再循环内体。此外，TEM 分析显示，在 90 分钟内，经历二次包膜的衣壳中有近 90%和 50%的包膜衣壳是 WGA 阳性的。我们的数据揭示了在 HCMV 感染后期内吞区室的广泛重塑，其中内吞区室为病毒形态发生提供了优化的环境，并作为二次包膜的主要膜源。此外，我们表明二次包膜是一个快速过程，内吞的膜在几分钟内从质膜运输到 cVAC，以供衣壳用于包膜。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/c1f009105b93/biomolecules-14-01149-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/6653b40dbfc2/biomolecules-14-01149-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/5d4a99395991/biomolecules-14-01149-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/c1a9c49b743a/biomolecules-14-01149-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/6832c41a0df9/biomolecules-14-01149-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/3abb6edb926f/biomolecules-14-01149-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/e05f25557fe0/biomolecules-14-01149-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/2193b758fb74/biomolecules-14-01149-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/239191baabba/biomolecules-14-01149-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/6cd33615c7aa/biomolecules-14-01149-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/c1f009105b93/biomolecules-14-01149-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/6653b40dbfc2/biomolecules-14-01149-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/5d4a99395991/biomolecules-14-01149-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/c1a9c49b743a/biomolecules-14-01149-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/6832c41a0df9/biomolecules-14-01149-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/3abb6edb926f/biomolecules-14-01149-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/e05f25557fe0/biomolecules-14-01149-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/2193b758fb74/biomolecules-14-01149-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/239191baabba/biomolecules-14-01149-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/6cd33615c7aa/biomolecules-14-01149-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5012/11430300/c1f009105b93/biomolecules-14-01149-g010.jpg

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