PMID- 31905776 OWN - NLM STAT- MEDLINE DCOM- 20200928 LR - 20200928 IS - 1999-4915 (Electronic) IS - 1999-4915 (Linking) VI - 12 IP - 1 DP - 2019 Dec 29 TI - MYH9 Key Amino Acid Residues Identified by the Anti-Idiotypic Antibody to Porcine Reproductive and Respiratory Syndrome Virus Glycoprotein 5 Involve in the Virus Internalization by Porcine Alveolar Macrophages. LID - 10.3390/v12010040 [doi] LID - 40 AB - MYH9 has been identified as an indispensable cellular protein for porcine reproductive and respiratory syndrome virus (PRRSV) entry into permissive cells using the monoclonal anti-idiotypic antibody (Mab2-5G2) recognizing an antibody that specifically interacts with PRRSV glycoprotein 5 (GP5). More recently, we found that Mab2-5G2 interacted with the MYH9 C-terminal domain, designated PRA, which is required for PRRSV internalization. In this study, we demonstrate that blocking of MYH9 with Mab2-5G2 significantly diminished PRRSV internalization by porcine alveolar macrophage (PAM) via interruption of direct interaction between GP5 and MYH9, and thus remarkably inhibited subsequent infection of PAMs by PRRSV-2 isolates. Moreover, the three-dimensional structure of the Mab2-5G2 Fab-PRA complex determined via homology modeling predicted potential docking sites required for PRRSV internalization. Further analysis of Mab2-5G2-binding sites within PRA highlighted that the amino acids E1670, K1673, E1679, and I1683 in PRA are the key Mab2-5G2-binding residues. Notably, recombinant PRA protein blocked the interaction between PRRSV GP5 and cellular MYH9 by preventing translocation of MYH9 from the cytoplasm to the cell membrane, an essential step for PRRSV virion internalization. Meanwhile, porcine cell line permissive for PRRSV bearing point mutation of E1670A in MYH9 demonstrated reduced susceptibility for PRRSV infection. In conclusion, this work increases understanding of both PRRSV pathogenesis and the mechanistic role played by MYH9 in PRRSV infection. FAU - Li, Liangliang AU - Li L AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. AD - College of Agronomy, Liaocheng University, Liaocheng 252059, China. FAU - Zhang, Lu AU - Zhang L AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Hu, Qifan AU - Hu Q AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Zhao, Liang AU - Zhao L AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Nan, Yuchen AU - Nan Y AUID- ORCID: 0000-0002-4442-8004 AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Hou, Gaopeng AU - Hou G AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Chen, Yiyang AU - Chen Y AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Han, Ximeng AU - Han X AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Ren, Xiaolei AU - Ren X AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Zhao, Qin AU - Zhao Q AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Tao, Hu AU - Tao H AUID- ORCID: 0000-0002-7347-2362 AD - College of Life Science, Northwest A&F University, Yangling 712100, China. FAU - Sun, Zhenzhao AU - Sun Z AUID- ORCID: 0000-0002-3936-9791 AD - Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 100193, China. FAU - Zhang, Gaiping AU - Zhang G AD - College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China. FAU - Wu, Chunyan AU - Wu C AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. FAU - Wang, Jingfei AU - Wang J AD - Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 100193, China. FAU - Zhou, En-Min AU - Zhou EM AD - Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20191229 PL - Switzerland TA - Viruses JT - Viruses JID - 101509722 RN - 0 (Amino Acids) RN - 0 (Antibodies, Anti-Idiotypic) RN - 0 (Antibodies, Monoclonal) RN - 0 (Viral Envelope Proteins) RN - 0 (glycoprotein 5, PRRSV) RN - EC 3.6.4.1 (Myosin Heavy Chains) SB - IM MH - Amino Acids/chemistry MH - Animals MH - Antibodies, Anti-Idiotypic/*immunology MH - Antibodies, Monoclonal/immunology MH - Binding Sites, Antibody MH - Macrophages, Alveolar/*immunology/*virology MH - Myosin Heavy Chains/*immunology MH - Porcine Reproductive and Respiratory Syndrome/virology MH - Porcine respiratory and reproductive syndrome virus/*immunology/physiology MH - Swine MH - Viral Envelope Proteins/*immunology MH - *Virus Internalization PMC - PMC7019770 OTO - NOTNLM OT - MYH9 OT - PRRSV OT - PRRSV GP5 OT - anti-idiotype OT - pathogenesis COIS- The authors declare no conflict of interest. EDAT- 2020/01/08 06:00 MHDA- 2020/09/29 06:00 PMCR- 2020/01/01 CRDT- 2020/01/08 06:00 PHST- 2019/12/15 00:00 [received] PHST- 2019/12/24 00:00 [accepted] PHST- 2020/01/08 06:00 [entrez] PHST- 2020/01/08 06:00 [pubmed] PHST- 2020/09/29 06:00 [medline] PHST- 2020/01/01 00:00 [pmc-release] AID - v12010040 [pii] AID - viruses-12-00040 [pii] AID - 10.3390/v12010040 [doi] PST - epublish SO - Viruses. 2019 Dec 29;12(1):40. doi: 10.3390/v12010040.