[Screening of cellular proteins binding to the core region of hepatitis C virus RNA by ultraviolet cross-linking assay].
OBJECTIVE
To screen cellular proteins binding to the core region of hepatitis C virus (HCV) from human hepatoma cells.


METHODS
Unlabeled 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.


RESULTS
Two 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.


CONCLUSIONS
The cellular proteins from HepG2 cells could bind to the core region of HCV RNA.
Su
Hai-xia
HX
Department of Epidemiology, Faculty of Preventive Medicine, Fourth Military Medical University, Xi'an 710033, China.
Zhang
Jing-xia
JX
Zhao
Xiao-ning
XN
Lu
Juan
J
Yan
Yong-ping
YP
D001665:Binding Sites; D003432:Cross-Linking Reagents / Q000737:chemistry; D016174:Hepacivirus / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D014466:Ultraviolet Rays; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*
D003432:Cross-Linking Reagents; D012367:RNA, Viral; D014758:Viral Core Proteins