title: Heat shock protein 70 is associated with CSFV NS5A protein and enhances viral RNA replication.
abstract: The non-structural 5A (NS5A) protein of classical swine fever virus (CSFV) is proven to be involved in viral replication and can also modulate cellular signaling via to its ability to interact with various cellular proteins. Here, HSP70/NS5A complex formation is confirmed by coimmunoprecipitation and GST-pulldown studies. Additionally, the N-terminal amino acids (29-240) of NS5A were identified as the interaction region through in vivo deletion analyses, and confocal microscopy showed that NS5A and HSP70 colocalized in the cytoplasm. Overexpression of HSP70 via the eukaryotic expression plasmid pDsRED N1 or lentivirus significantly promoted viral RNA synthesis. Whereas the knockdown of HSP70 by lentivirus-mediated shRNA or inhibition by quercetin markedly decreased the viral load. These data suggest that HSP70 plays a critical role in the viral life cycle, particularly during the virus RNA replication period. The investigation of HSP70 protein functions may be beneficial for developing new strategies to treat CSFV infection.
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Chengcheng
authors.initials: C
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Kai
authors.initials: K
authors.lastname: Kang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Pengbo
authors.initials: P
authors.lastname: Ning
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Yangxin
authors.initials: Y
authors.lastname: Peng
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Lin
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Hongjie
authors.initials: H
authors.lastname: Cui
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Cao
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi, PR China. Electronic address: zhangym@nwsuaf.edu.cn.
authors.forename: Yanming
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D002498:Centrifugation; D006692:Classical Swine Fever Virus / Q000502:physiology*; D004252:DNA Mutational Analysis; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D047468:Immunoprecipitation; D018613:Microscopy, Confocal; D011485:Protein Binding; D025941:Protein Interaction Mapping; D012367:RNA, Viral / Q000378:metabolism*; D017384:Sequence Deletion; D017361:Viral Nonstructural Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D018840:HSP70 Heat-Shock Proteins; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins
keywords: Classical swine fever virus (CSFV); HSP70; NS5A