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[Determination of DNA binding domains in hepatitis C virus core protein]
Objective:
To determine the DNA binding domain in hepatitis C virus core protein and otelucidate the significance of binding.
Methods:
Segments of hepatitis C virus core protein were expressed in E.coli as fusion forms with glutathion S-transferase (GST). The core proteins were immobilized in SDS-PAGE gel after removing SDS from the gel by washing. (32)P-ATP labeled oligonucleotides were electrophoresed through the gel in TAE buffer. The binding of DNA with core protein was detected by autoradiography.
Results:
There were at least two DNA binding domains in the HCV core protein, the first locating at 10~16aa and the second at 46-70aa. The second region was divided into three adjacent parts, which could bind core protein independently. Core protein bound to single strand DNA as well as to double strand DNA without sequence specificity.
Conclusions:
DNA binding regions of HCV core protein locate at its N-terminus. The binding regions of HCV core protein overlap its nucleus transfer signals and they bind to target DNA unselectively, suggesting a possible mechanism for its multifunction. The result provides basic data for understanding the biological function of HCV core protein.