Screening of cellular proteins binding to the core region of hepatitis C virus RNA by ultraviolet cross-linking assay.
- Author:
Hai-xia SU
1
;
Jing-xia ZHANG
;
Xiao-ning ZHAO
;
Juan LU
;
Yong-ping YAN
Author Information
- Publication Type:Journal Article
- MeSH: Binding Sites; Cross-Linking Reagents; chemistry; Hepacivirus; genetics; metabolism; RNA, Viral; genetics; metabolism; Ultraviolet Rays; Viral Core Proteins; genetics; metabolism
- From: Chinese Journal of Hepatology 2005;13(9):656-659
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screen cellular proteins binding to the core region of hepatitis C virus (HCV) from human hepatoma cells.
METHODSUnlabeled and labeled RNA transcripts were prepared by in vitro transcription. Cytoplasmic extracts were prepared from human hepatoma cells HepG2. Ultraviolet (UV) cross-linking was used to screen the cellular proteins that would bind to the core region of HCV. Competition experiment was performed to confirm the specificity of the binding in which excess unlabeled RNA of HCV core region and plasmid RNA were used as competitors.
RESULTSTwo cellular proteins of 6.6 x 10(4) and 5.5 x 10(4) were found binding to the core region of HCV RNA by UV cross-linking assay. The unlabeled core region of HCV RNA could compete out this binding whereas the unlabeled plasmid RNA could not.
CONCLUSIONThe cellular proteins from HepG2 cells could bind to the core region of HCV RNA.