A C-terminal truncated hepatitis C virus core protein variant assembles in vitro into virus-like particles in the absence of structured nucleic acids.
Little is known about the assembly pathway or structure of the hepatitis C virus (HCV). In this work a truncated HCcAg variant covering the first 120 aa (HCcAg.120) with a 32 aa N-terminal fusion peptide (6x Histag-Xpress epitope) was purified as a monomer under strong denaturing conditions. In addition, minor HCcAg.120 peaks exhibiting little different molecular mass by SDS-PAGE which possibly represents alternative forms harboring the N-termini of HCcAg.120 were detected. Analysis using gel filtration chromatography showed that HCcAg.120 assembled into high molecular weight structures in vitro in the absence of structured nucleic acids. The negative-stain electron microscopy analysis revealed that these structures correspond with spherical VLPs of uniform morphology and size distribution. The diameters of these particles ranged from 20 to 43nm with an average diameter of approximately 30 nm and were specifically immunolabelled with a mouse monoclonal antibody against the residues 5-35 of HCcAg. Results presented in this work showed that HCcAg.120 assembled in vitro into VLPs in the absence of structured nucleic acids with similar morphology and size distribution to those found in sera and hepatocytes from HCV-infected patients. Therefore, these VLPs would be important to elucidate the mechanisms behind the ability of HCcAg to assemble into a nucleocapsid structure.
Acosta-Rivero
Nelson
N
Biomedical Research Division, Center for Genetic Engineering and Biotechnology, P.O. Box 6162, C.P. 10600, C. Habana, Cuba. nelson.acosta@cigb.edu.cu
Rodriguez
Armando
A
Mussachio
Alexis
A
Poutou
Johana
J
Falcon
Viviana
V
Torres
Dinorah
D
Aguilar
Julio C
JC
Linares
Marbelis
M
Alonso
Mabel
M
Perez
Angel
A
Menendez
Ivon
I
Morales-Grillo
Juan
J
Marquez
Gabriel
G
Dueñas-Carrera
Santiago
S
D016174:Hepacivirus / Q000378:metabolism* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D019251:Nucleocapsid / Q000096:biosynthesis*; D010446:Peptide Fragments / Q000378:metabolism*; D012367:RNA, Viral; D014758:Viral Core Proteins / Q000378:metabolism*
D010446:Peptide Fragments; D012367:RNA, Viral; D014758:Viral Core Proteins; C511446:core protein (1-120), hepatitis C virus