Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive Cells
Porcine reproductive and respiratory syndrome virus (PRRSV) has a highly restricted tropism for cells of the monocyte-macrophage lineage, including porcine alveolar macrophages (PAMs). PRRSV entry into permissive cells involves several mediators in addition to two required host cell receptors, CD163...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2019-08-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fmicb.2019.01815/full |
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| author | Gaopeng Hou Biyun Xue Liangliang Li Yuchen Nan Lu Zhang Kuokuo Li Qin Zhao Julian A. Hiscox James P. Stewart Chunyan Wu Jingfei Wang En-Min Zhou |
| author_facet | Gaopeng Hou Biyun Xue Liangliang Li Yuchen Nan Lu Zhang Kuokuo Li Qin Zhao Julian A. Hiscox James P. Stewart Chunyan Wu Jingfei Wang En-Min Zhou |
| author_sort | Gaopeng Hou |
| collection | DOAJ |
| description | Porcine reproductive and respiratory syndrome virus (PRRSV) has a highly restricted tropism for cells of the monocyte-macrophage lineage, including porcine alveolar macrophages (PAMs). PRRSV entry into permissive cells involves several mediators in addition to two required host cell receptors, CD163 and MYH9. It is unknown whether CD163 directly interacts and/or cooperates with MYH9 to facilitate PRRSV infection. In this study, CD163 and MYH9 were co-immunoprecipitated from PAMs regardless of PRRSV infection status. Further truncation analysis indicated that the CD163 N-terminal region, containing scavenger receptor cysteine-rich domains 1 to 4 (SRCR1-4), directly interacts with the MYH9 C-terminal domain region without involvement of other adaptor proteins. Meanwhile, non-permissive HEK293T cells that stably expressed truncated swine CD163 SRCR1-4 domain did not support virus attachment. However, virus attachment to cells stably expressing SRCR5-CT domain was demonstrated to occur without appreciable virus internalization. The involvement of the SRCR1-4 domain in virus internalization was further demonstrated by the fact that incubation of recombinant SRCR1-4 protein with PAMs abolished subsequent virus internalization by permissive cells. These results demonstrated that CD163 SRCR1-4 interacts with the MYH9 C–terminal domain to facilitate PRRSV virion internalization in permissive cells, thus expanding our understanding of PRRSV cell-invasion mechanisms. |
| first_indexed | 2024-12-11T18:12:45Z |
| format | Article |
| id | doaj.art-83cf9ff2ff754eacb8a7c94f93b763e4 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-11T18:12:45Z |
| publishDate | 2019-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-83cf9ff2ff754eacb8a7c94f93b763e42022-12-22T00:55:32ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2019-08-011010.3389/fmicb.2019.01815471634Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive CellsGaopeng Hou0Biyun Xue1Liangliang Li2Yuchen Nan3Lu Zhang4Kuokuo Li5Qin Zhao6Julian A. Hiscox7James P. Stewart8Chunyan Wu9Jingfei Wang10En-Min Zhou11Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaDepartment of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool, United KingdomDepartment of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool, United KingdomDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaDepartment of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling, ChinaPorcine reproductive and respiratory syndrome virus (PRRSV) has a highly restricted tropism for cells of the monocyte-macrophage lineage, including porcine alveolar macrophages (PAMs). PRRSV entry into permissive cells involves several mediators in addition to two required host cell receptors, CD163 and MYH9. It is unknown whether CD163 directly interacts and/or cooperates with MYH9 to facilitate PRRSV infection. In this study, CD163 and MYH9 were co-immunoprecipitated from PAMs regardless of PRRSV infection status. Further truncation analysis indicated that the CD163 N-terminal region, containing scavenger receptor cysteine-rich domains 1 to 4 (SRCR1-4), directly interacts with the MYH9 C-terminal domain region without involvement of other adaptor proteins. Meanwhile, non-permissive HEK293T cells that stably expressed truncated swine CD163 SRCR1-4 domain did not support virus attachment. However, virus attachment to cells stably expressing SRCR5-CT domain was demonstrated to occur without appreciable virus internalization. The involvement of the SRCR1-4 domain in virus internalization was further demonstrated by the fact that incubation of recombinant SRCR1-4 protein with PAMs abolished subsequent virus internalization by permissive cells. These results demonstrated that CD163 SRCR1-4 interacts with the MYH9 C–terminal domain to facilitate PRRSV virion internalization in permissive cells, thus expanding our understanding of PRRSV cell-invasion mechanisms.https://www.frontiersin.org/article/10.3389/fmicb.2019.01815/fullPRRSVCD163MYH9protein–protein interactionvirus internalization |
| spellingShingle | Gaopeng Hou Biyun Xue Liangliang Li Yuchen Nan Lu Zhang Kuokuo Li Qin Zhao Julian A. Hiscox James P. Stewart Chunyan Wu Jingfei Wang En-Min Zhou Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive Cells Frontiers in Microbiology PRRSV CD163 MYH9 protein–protein interaction virus internalization |
| title | Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive Cells |
| title_full | Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive Cells |
| title_fullStr | Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive Cells |
| title_full_unstemmed | Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive Cells |
| title_short | Direct Interaction Between CD163 N-Terminal Domain and MYH9 C-Terminal Domain Contributes to Porcine Reproductive and Respiratory Syndrome Virus Internalization by Permissive Cells |
| title_sort | direct interaction between cd163 n terminal domain and myh9 c terminal domain contributes to porcine reproductive and respiratory syndrome virus internalization by permissive cells |
| topic | PRRSV CD163 MYH9 protein–protein interaction virus internalization |
| url | https://www.frontiersin.org/article/10.3389/fmicb.2019.01815/full |
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