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. CONCLUSION:PGAM-B could specifically bind to the core region of HCV RNA genome in vitro.
journal_name
Intervirologyjournal_title
Intervirologyauthors
Su HX,Xu DZ,Zhang YH,Men K,Zhao XN,Li D,Zhang L,Zhang JX,Yan YPdoi
10.1159/000105443subject
Has Abstractpub_date
2007-01-01 00:00:00pages
303-9issue
4eissn
0300-5526issn
1423-0100pii
000105443journal_volume
50pub_type
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