Insights into the function of ESCRT complex and LBPA in ASFV infection
The African swine fever virus (ASFV) is strongly dependent on an intact endocytic pathway and a certain cellular membrane remodeling for infection, possibly regulated by the endosomal sorting complexes required for transport (ESCRT). The ESCRT machinery is mainly involved in the coordination of memb...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-12-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1163569/full |
| _version_ | 1797403355045167104 |
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| author | Lucía Barrado-Gil Isabel García-Dorival Inmaculada Galindo Covadonga Alonso Miguel Ángel Cuesta-Geijo |
| author_facet | Lucía Barrado-Gil Isabel García-Dorival Inmaculada Galindo Covadonga Alonso Miguel Ángel Cuesta-Geijo |
| author_sort | Lucía Barrado-Gil |
| collection | DOAJ |
| description | The African swine fever virus (ASFV) is strongly dependent on an intact endocytic pathway and a certain cellular membrane remodeling for infection, possibly regulated by the endosomal sorting complexes required for transport (ESCRT). The ESCRT machinery is mainly involved in the coordination of membrane dynamics; hence, several viruses exploit this complex and its accessory proteins VPS4 and ALIX for their own benefit. In this work, we found that shRNA-mediated knockdown of VPS4A decreased ASFV replication and viral titers, and this silencing resulted in an enhanced expression of ESCRT-0 component HRS. ASFV infection slightly increased HRS expression but not under VPS4A depletion conditions. Interestingly, VPS4A silencing did not have an impact on ALIX expression, which was significantly overexpressed upon ASFV infection. Further analysis revealed that ALIX silencing impaired ASFV infection at late stages of the viral cycle, including replication and viral production. In addition to ESCRT, the accessory protein ALIX is involved in endosomal membrane dynamics in a lysobisphosphatydic acid (LBPA) and Ca2+-dependent manner, which is relevant for intraluminal vesicle (ILV) biogenesis and endosomal homeostasis. Moreover, LBPA interacts with NPC2 and/or ALIX to regulate cellular cholesterol traffic, and would affect ASFV infection. Thus, we show that LBPA blocking impacted ASFV infection at both early and late infection, suggesting a function for this unconventional phospholipid in the ASFV viral cycle. Here, we found for the first time that silencing of VPS4A and ALIX affects the infection later on, and blocking LBPA function reduces ASFV infectivity at early and later stages of the viral cycle, while ALIX was overexpressed upon infection. These data suggested the relevance of ESCRT-related proteins in ASFV infection. |
| first_indexed | 2024-03-09T02:37:34Z |
| format | Article |
| id | doaj.art-8b2e9f316ee34aac950aac70aca1c9cf |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-03-09T02:37:34Z |
| publishDate | 2023-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-8b2e9f316ee34aac950aac70aca1c9cf2023-12-06T08:25:25ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882023-12-011310.3389/fcimb.2023.11635691163569Insights into the function of ESCRT complex and LBPA in ASFV infectionLucía Barrado-GilIsabel García-DorivalInmaculada GalindoCovadonga AlonsoMiguel Ángel Cuesta-GeijoThe African swine fever virus (ASFV) is strongly dependent on an intact endocytic pathway and a certain cellular membrane remodeling for infection, possibly regulated by the endosomal sorting complexes required for transport (ESCRT). The ESCRT machinery is mainly involved in the coordination of membrane dynamics; hence, several viruses exploit this complex and its accessory proteins VPS4 and ALIX for their own benefit. In this work, we found that shRNA-mediated knockdown of VPS4A decreased ASFV replication and viral titers, and this silencing resulted in an enhanced expression of ESCRT-0 component HRS. ASFV infection slightly increased HRS expression but not under VPS4A depletion conditions. Interestingly, VPS4A silencing did not have an impact on ALIX expression, which was significantly overexpressed upon ASFV infection. Further analysis revealed that ALIX silencing impaired ASFV infection at late stages of the viral cycle, including replication and viral production. In addition to ESCRT, the accessory protein ALIX is involved in endosomal membrane dynamics in a lysobisphosphatydic acid (LBPA) and Ca2+-dependent manner, which is relevant for intraluminal vesicle (ILV) biogenesis and endosomal homeostasis. Moreover, LBPA interacts with NPC2 and/or ALIX to regulate cellular cholesterol traffic, and would affect ASFV infection. Thus, we show that LBPA blocking impacted ASFV infection at both early and late infection, suggesting a function for this unconventional phospholipid in the ASFV viral cycle. Here, we found for the first time that silencing of VPS4A and ALIX affects the infection later on, and blocking LBPA function reduces ASFV infectivity at early and later stages of the viral cycle, while ALIX was overexpressed upon infection. These data suggested the relevance of ESCRT-related proteins in ASFV infection.https://www.frontiersin.org/articles/10.3389/fcimb.2023.1163569/fullESCRTLBPAAlixVps4multivesicular bodiesNPC1 |
| spellingShingle | Lucía Barrado-Gil Isabel García-Dorival Inmaculada Galindo Covadonga Alonso Miguel Ángel Cuesta-Geijo Insights into the function of ESCRT complex and LBPA in ASFV infection Frontiers in Cellular and Infection Microbiology ESCRT LBPA Alix Vps4 multivesicular bodies NPC1 |
| title | Insights into the function of ESCRT complex and LBPA in ASFV infection |
| title_full | Insights into the function of ESCRT complex and LBPA in ASFV infection |
| title_fullStr | Insights into the function of ESCRT complex and LBPA in ASFV infection |
| title_full_unstemmed | Insights into the function of ESCRT complex and LBPA in ASFV infection |
| title_short | Insights into the function of ESCRT complex and LBPA in ASFV infection |
| title_sort | insights into the function of escrt complex and lbpa in asfv infection |
| topic | ESCRT LBPA Alix Vps4 multivesicular bodies NPC1 |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1163569/full |
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