Structural requirements for assembly and homotypic interactions of the hepatitis C virus core protein.
The hepatitis C virus (HCV) core protein is involved in the assembly of nucleocapsid particles, as well as regulation of cellular and viral gene expression. To investigate the biological properties of the viral core protein and viral RNA assembly, two recombinant core proteins, the mature core protein (named C179) and a C-terminal truncated protein (named C124), were expressed and purified. To confirm their ability to generate viral particles, the production of nucleocapsid-like particles was monitored using transmission electron microscopy (EM). The EM analysis revealed that exposure of these proteins to the 5' untranslated region (5' UTR) of the viral RNA resulted in generation of spherical particles of 30-140nm in diameter. Interestingly, a cross-linking analysis revealed that C124 required an RNA component for homotypic interactions. In contrast, C179 successfully assembled in the absence of nucleic acids. Additionally, RNA-mediated conversion of the C124 structure into a more stable state was maintained even after RNase treatment. Therefore, our results indicate that the basic N-terminal domain of the viral core protein utilizes RNA components to induce conformational changes or efficient homotypic interactions, while the C-terminal domain may contain key peptide sequences for initiating spontaneous multimerization at the early stages of viral assembly.
Kim
Meehyein
M
Immunology and Virology Group, Mogam Biotechnology Research Institute, 341, Pojung-ri, Guseong-eup, Yongin-city, Kyonggi-do 449-913, South Korea. mkin@morgan.re.kr
Ha
Youngju
Y
Park
Hae-Joon
HJ
D020121:5' Untranslated Regions / Q000378:metabolism; D003001:Cloning, Molecular; D016174:Hepacivirus / Q000502:physiology*; D046529:Microscopy, Electron, Transmission; D019251:Nucleocapsid / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure; D012367:RNA, Viral / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014758:Viral Core Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D019065:Virus Assembly
D020121:5' Untranslated Regions; D012367:RNA, Viral; D011994:Recombinant Proteins; D014758:Viral Core Proteins; C069130:nucleocapsid protein, Hepatitis C virus