title: Interaction of structural core protein of classical swine fever virus with endoplasmic reticulum-associated degradation pathway protein OS9.
abstract: Classical swine fever virus (CSFV) Core protein is involved in virus RNA protection, transcription regulation and virus virulence. To discover additional Core protein functions a yeast two-hybrid system was used to identify host proteins that interact with Core. Among the identified host proteins, the osteosarcoma amplified 9 protein (OS9) was further studied. Using alanine scanning mutagenesis, the OS9 binding site in the CSFV Core protein was identified, between Core residues (90)IAIM(93), near a putative cleavage site. Truncated versions of Core were used to show that OS9 binds a polypeptide representing the 12 C-terminal Core residues. Cells transfected with a double-fluorescent labeled Core construct demonstrated that co-localization of OS9 and Core occurred only on unprocessed forms of Core protein. A recombinant CSFV containing Core protein where residues (90)IAIM(93) were substituted by alanines showed no altered virulence in swine, but a significant decreased ability to replicate in cell cultures.
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: douglas.gladue@ars.usda.gov.
authors.forename: D P
authors.initials: DP
authors.lastname: Gladue
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: vivian.odonnell@ars.usda.gov.
authors.forename: V
authors.initials: V
authors.lastname: O'Donnell
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: ignacio.fernandez-sainz@ars.usda.gov.
authors.forename: I J
authors.initials: IJ
authors.lastname: Fernandez-Sainz
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: Paige.Fletcher@ars.usda.gov.
authors.forename: P
authors.initials: P
authors.lastname: Fletcher
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA; Plum Island Animal Disease Center, DHS, Greenport, NY 11944, USA. Electronic address: Ryan.Baker-Branstetter@ars.usda.gov.
authors.forename: R
authors.initials: R
authors.lastname: Baker-Branstetter
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: lauren.holinka@ars.usda.gov.
authors.forename: L G
authors.initials: LG
authors.lastname: Holinka
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: Brenton.Stanford@ars.usda.gov.
authors.forename: B
authors.initials: B
authors.lastname: Sanford
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: Jolene.Carlson@ars.usda.gov.
authors.forename: J
authors.initials: J
authors.lastname: Carlson
authors.affiliation: Plum Island Animal Disease Center, DHS, Greenport, NY 11944, USA. Electronic address: zlu@dhs.gov.
authors.forename: Z
authors.initials: Z
authors.lastname: Lu
authors.affiliation: Plum Island Animal Disease Center, ARS, USDA, Greenport, NY 11944, USA. Electronic address: manuel.borca@ars.usda.gov.
authors.forename: M V
authors.initials: MV
authors.lastname: Borca
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D006691:Classical Swine Fever / Q000235:genetics / Q000378:metabolism* / Q000821:virology; D006692:Classical Swine Fever Virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D060746:Endoplasmic Reticulum-Associated Degradation; D054884:Host-Pathogen Interactions; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D013552:Swine; D020798:Two-Hybrid System Techniques; D014758:Viral Core Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
chemical_list: D009363:Neoplasm Proteins; D014758:Viral Core Proteins
keywords: Attenuation; Classical swine fever virus; Core protein; Pathogenesis; Virulence