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Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor
The genus Sapovirus belongs to the family Caliciviridae, and its members are common causative agents of severe acute gastroenteritis in both humans and animals. Some caliciviruses are known to use either terminal sialic acids or histo-blood group antigens as attachment factors and/or cell surface pr...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6364031/ https://www.ncbi.nlm.nih.gov/pubmed/30463963 http://dx.doi.org/10.1128/JVI.01773-18 |
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author | Alfajaro, Mia Madel Cho, Eun-Hyo Kim, Deok-Song Kim, Ji-Yun Park, Jun-Gyu Soliman, Mahmoud Baek, Yeong-Bin Park, Chul-Ho Kang, Mun-Il Park, Sang-Ik Cho, Kyoung-Oh |
author_facet | Alfajaro, Mia Madel Cho, Eun-Hyo Kim, Deok-Song Kim, Ji-Yun Park, Jun-Gyu Soliman, Mahmoud Baek, Yeong-Bin Park, Chul-Ho Kang, Mun-Il Park, Sang-Ik Cho, Kyoung-Oh |
author_sort | Alfajaro, Mia Madel |
collection | PubMed |
description | The genus Sapovirus belongs to the family Caliciviridae, and its members are common causative agents of severe acute gastroenteritis in both humans and animals. Some caliciviruses are known to use either terminal sialic acids or histo-blood group antigens as attachment factors and/or cell surface proteins, such as CD300lf, CD300ld, and junctional adhesion molecule 1 of tight junctions (TJs), as receptors. However, the roles of TJs and their proteins in sapovirus entry have not been examined. In this study, we found that porcine sapovirus (PSaV) significantly decreased transepithelial electrical resistance and increased paracellular permeability early in infection of LLC-PK cells, suggesting that PSaV dissociates TJs of cells. This led to the interaction between PSaV particles and occludin, which traveled in a complex into late endosomes via Rab5- and Rab7-dependent trafficking. Inhibition of occludin using small interfering RNA (siRNA), a specific antibody, or a dominant-negative mutant significantly blocked the entry of PSaV. Transient expression of occludin in nonpermissive Chinese hamster ovary (CHO) cells conferred susceptibility to PSaV, but only for a limited time. Although claudin-1, another TJ protein, neither directly interacted nor was internalized with PSaV particles, it facilitated PSaV entry and replication in the LLC-PK cells. We conclude that PSaV particles enter LLC-PK cells by binding to occludin as a coreceptor in PSaV-dissociated TJs. PSaV and occludin then form a complex that moves to late endosomes via Rab5- and Rab7-dependent trafficking. In addition, claudin-1 in the TJs opened by PSaV infection facilitates PSaV entry and infection as an entry factor. IMPORTANCE Sapoviruses (SaVs) cause severe acute gastroenteritis in humans and animals. Although they replicate in intestinal epithelial cells, which are tightly sealed by apical-junctional complexes, such as tight junctions (TJs), the mechanisms by which SaVs hijack TJs and their proteins for successful entry and infection remain largely unknown. Here, we demonstrate that porcine SaVs (PSaVs) induce early dissociation of TJs, allowing them to bind to the TJ protein occludin as a functional coreceptor. PSaVs then travel in a complex with occludin into late endosomes through Rab5- and Rab7-dependent trafficking. Claudin-1, another TJ protein, does not directly interact with PSaV but facilitates the entry of PSaV into cells as an entry factor. This work contributes to our understanding of the entry of SaV and other caliciviruses into cells and may aid in the development of efficient and affordable drugs to treat SaV infections. |
format | Online Article Text |
id | pubmed-6364031 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-63640312019-02-22 Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor Alfajaro, Mia Madel Cho, Eun-Hyo Kim, Deok-Song Kim, Ji-Yun Park, Jun-Gyu Soliman, Mahmoud Baek, Yeong-Bin Park, Chul-Ho Kang, Mun-Il Park, Sang-Ik Cho, Kyoung-Oh J Virol Virus-Cell Interactions The genus Sapovirus belongs to the family Caliciviridae, and its members are common causative agents of severe acute gastroenteritis in both humans and animals. Some caliciviruses are known to use either terminal sialic acids or histo-blood group antigens as attachment factors and/or cell surface proteins, such as CD300lf, CD300ld, and junctional adhesion molecule 1 of tight junctions (TJs), as receptors. However, the roles of TJs and their proteins in sapovirus entry have not been examined. In this study, we found that porcine sapovirus (PSaV) significantly decreased transepithelial electrical resistance and increased paracellular permeability early in infection of LLC-PK cells, suggesting that PSaV dissociates TJs of cells. This led to the interaction between PSaV particles and occludin, which traveled in a complex into late endosomes via Rab5- and Rab7-dependent trafficking. Inhibition of occludin using small interfering RNA (siRNA), a specific antibody, or a dominant-negative mutant significantly blocked the entry of PSaV. Transient expression of occludin in nonpermissive Chinese hamster ovary (CHO) cells conferred susceptibility to PSaV, but only for a limited time. Although claudin-1, another TJ protein, neither directly interacted nor was internalized with PSaV particles, it facilitated PSaV entry and replication in the LLC-PK cells. We conclude that PSaV particles enter LLC-PK cells by binding to occludin as a coreceptor in PSaV-dissociated TJs. PSaV and occludin then form a complex that moves to late endosomes via Rab5- and Rab7-dependent trafficking. In addition, claudin-1 in the TJs opened by PSaV infection facilitates PSaV entry and infection as an entry factor. IMPORTANCE Sapoviruses (SaVs) cause severe acute gastroenteritis in humans and animals. Although they replicate in intestinal epithelial cells, which are tightly sealed by apical-junctional complexes, such as tight junctions (TJs), the mechanisms by which SaVs hijack TJs and their proteins for successful entry and infection remain largely unknown. Here, we demonstrate that porcine SaVs (PSaVs) induce early dissociation of TJs, allowing them to bind to the TJ protein occludin as a functional coreceptor. PSaVs then travel in a complex with occludin into late endosomes through Rab5- and Rab7-dependent trafficking. Claudin-1, another TJ protein, does not directly interact with PSaV but facilitates the entry of PSaV into cells as an entry factor. This work contributes to our understanding of the entry of SaV and other caliciviruses into cells and may aid in the development of efficient and affordable drugs to treat SaV infections. American Society for Microbiology 2019-02-05 /pmc/articles/PMC6364031/ /pubmed/30463963 http://dx.doi.org/10.1128/JVI.01773-18 Text en Copyright © 2019 Alfajaro et al. https://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Virus-Cell Interactions Alfajaro, Mia Madel Cho, Eun-Hyo Kim, Deok-Song Kim, Ji-Yun Park, Jun-Gyu Soliman, Mahmoud Baek, Yeong-Bin Park, Chul-Ho Kang, Mun-Il Park, Sang-Ik Cho, Kyoung-Oh Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor |
title | Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor |
title_full | Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor |
title_fullStr | Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor |
title_full_unstemmed | Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor |
title_short | Early Porcine Sapovirus Infection Disrupts Tight Junctions and Uses Occludin as a Coreceptor |
title_sort | early porcine sapovirus infection disrupts tight junctions and uses occludin as a coreceptor |
topic | Virus-Cell Interactions |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6364031/ https://www.ncbi.nlm.nih.gov/pubmed/30463963 http://dx.doi.org/10.1128/JVI.01773-18 |
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