title: Screening cellular proteins binding to the core region of hepatitis C virus RNA genome with digoxin-labeled nucleic acids.
abstract: OBJECTIVE
  To screen and identify cellular proteins binding to the core region of hepatitis C virus (HCV) RNA genome.
  
  
  METHODS
  The plasmid pHCV core was constructed to generate in vitro transcripts of the core region of HCV RNA genome. Ultraviolet (UV) cross-linking experiment and competition analysis were performed to screen HepG2 cellular proteins, which interact with digoxin-labeled transcripts of the core region of HCV RNA genome. RNA-binding proteins were separated by immunoprecipitation, analyzed by electrophoresis on SDS-PAGE and detected by immunoblotting with anti-digoxingenin-AP. After being excised from SDS-PAGE, the proteins bands were analyzed by MALDI-TOF-MS.
  
  
  RESULTS
  Several cellular proteins of hepG2 cell specifically bound to the core region of HCV RNA genome. The binding of cellular proteins to digoxin-labeled HCV core RNA was competed out in proportion to the increasing amount of unlabeled RNA. One of the HCV RNA-binding proteins was the B (brain) isozyme of human phosphoglycerate mutase (PGAM-B) identified by MALDI-TOF-MS.
  
  
  CONCLUSIONS
  PGAM-B could specifically bind to the core region of HCV RNA genome in vitro.
authors.affiliation: Department of Epidemiology, Fourth Military Medical University, Xi'an, China.
authors.forename: Hai-Xia
authors.initials: HX
authors.lastname: Su
authors.forename: De-Zhong
authors.initials: DZ
authors.lastname: Xu
authors.forename: Yu-Hai
authors.initials: YH
authors.lastname: Zhang
authors.forename: Ke
authors.initials: K
authors.lastname: Men
authors.forename: Xiao-Ning
authors.initials: XN
authors.lastname: Zhao
authors.forename: Duan
authors.initials: D
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Jing-Xia
authors.initials: JX
authors.lastname: Zhang
authors.forename: Yong-Ping
authors.initials: YP
authors.lastname: Yan
mesh_terms: D001665:Binding Sites; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000737:chemistry; D004077:Digoxin / Q000737:chemistry*; D016174:Hepacivirus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D010736:Phosphoglycerate Mutase / Q000378:metabolism*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism*; D014466:Ultraviolet Rays; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D003432:Cross-Linking Reagents; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D014758:Viral Core Proteins; D004077:Digoxin; D010736:Phosphoglycerate Mutase