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. CONCLUSION:PGAM-B could specifically bind to the core region of HCV RNA genome in vitro.

journal_name

Intervirology

journal_title

Intervirology

authors

Su HX,Xu DZ,Zhang YH,Men K,Zhao XN,Li D,Zhang L,Zhang JX,Yan YP

doi

10.1159/000105443

subject

Has Abstract

pub_date

2007-01-01 00:00:00

pages

303-9

issue

4

eissn

0300-5526

issn

1423-0100

pii

000105443

journal_volume

50

pub_type

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