Self-association of the C-terminal domain of the hepatitis-C virus core protein.
The N-terminal region of the hepatitis-C virus (HCV) core protein is rich in basic residues, while the C-terminal end of the protein comprises of a stretch of hydrophobic amino acids. Between these two extremes is an amphipathic region with two predicted alpha-helical segments. This region embodies Leu or hydrophobic residues in positions of heptad repeats and is possibly capable of self-association. To investigate this possibility, the core sequence was divided into two fragments and expressed separately as recombinant proteins. Recombinant proteins with the N-terminal fragment remained as monomers even at high concentrations in SDS/PAGE. Recombinant protein with the C-terminal fragment appeared largely monomeric on denaturing gels but some oligomers were also detected. Furthermore, proline mutations in either one of the predicted alpha helices adversely affected the observed oligomerization. The self-association capacity of the core protein C-terminal region was further supported by results from a yeast two-hybrid system. To affirm our conclusion, a peptide covering the heptad repeats and the predicted alpha helices was synthesized. Data from mass spectrometry and gel-filtration chromatography concluded that this peptide readily self-associated into the homodimer. Therefore, our results suggest that the oligomerization motifs of the HCV core protein may not be limited to the previously suggested N-terminal region.
Yan
B S
BS
Institute of Microbiology and Immunology, National Yang Ming University, Taipei, Taiwan.
Tam
M H
MH
Syu
W J
WJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D001704:Biopolymers; D017931:DNA Primers; D016174:Hepacivirus / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014758:Viral Core Proteins / Q000737:chemistry* / Q000235:genetics
D001704:Biopolymers; D017931:DNA Primers; D011994:Recombinant Proteins; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus