title: Identification of cleavage of NS5A of C-strain classical swine fever virus.
abstract: NS5A is a multifunctional non-structural protein of classical swine fever virus (CSFV) that plays an important role in viral replication, but how it exerts its functions is unknown. Here, we report the cleavage of NS5A of the vaccine C-strain, resulting in two truncated forms (b and c). Further experiments using calpain- and caspase-family-specific inhibitors, followed by a caspase-6-specific shRNAs and inhibitor, showed that the cleavage of C-strain NS5A to produce truncated form c is mediated by caspase-6, mapping to272DTTD275, while the cleavage producing truncated form b is probably mediated by another unknown protease. shRNA-mediated downregulation of caspase-6 and blocking of enzyme activity in ST cells significantly impaired genome replication and virus production, indicating that NS5A cleavage is required for CSFV replication.
authors.affiliation: College of Veterinary Medicine, Jilin University, Changchun, 130062, China.|Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun, 130122, China.
authors.forename: Jinxin
authors.initials: J
authors.lastname: Xie
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun, 130122, China.
authors.forename: Huancheng
authors.initials: H
authors.lastname: Guo
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun, 130122, China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Gong
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun, 130122, China.
authors.forename: Daliang
authors.initials: D
authors.lastname: Jiang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun, 130122, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun, 130122, China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Jia
authors.affiliation: Key Laboratory of Jilin Province for Zoonosis Prevention and Control, Institute of Military Veterinary, Academy of Military Medical Sciences, 666 Liuying West Road, Changchun, 130122, China. changchun_tu@hotmail.com.|Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, China. changchun_tu@hotmail.com.
authors.forename: Changchun
authors.initials: C
authors.lastname: Tu
mesh_terms: D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D002154:Calpain / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D053178:Caspase 6 / Q000235:genetics* / Q000378:metabolism; D061945:Caspase Inhibitors / Q000494:pharmacology; D002460:Cell Line; D006692:Classical Swine Fever Virus / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D007985:Leydig Cells / Q000187:drug effects* / Q000821:virology; D008297:Male; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D059748:Proteolysis / Q000187:drug effects; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013552:Swine; D013737:Testis / Q000187:drug effects / Q000821:virology; D017361:Viral Nonstructural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000590:Amino Acid Chloromethyl Ketones; D061945:Caspase Inhibitors; D011480:Protease Inhibitors; D034741:RNA, Small Interfering; D017361:Viral Nonstructural Proteins; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; D012324:RNA-Dependent RNA Polymerase; C505514:CASP6 protein, human; D002154:Calpain; D053178:Caspase 6