Severe acute respiratory syndrome coronavirus nucleocapsid protein confers ability to efficiently produce virus-like particles when substituted for the human immunodeficiency virus nucleocapsid domain.
We replaced the HIV-1 nucleocapsid (NC) domain with different N-coding sequences to test SARS-CoV nucleocapsid (N) self-interaction capacity, and determined the capabilities of each chimera to direct virus-like particle (VLP) assembly. Analysis results indicate that the replacement of NC with the carboxyl-terminal half of the SARS-CoV N resulted in the production of wild type (wt)-level virus-like particles (VLPs) with the density of a wt HIV-1 particle. When co-expressed with SARS-CoV N, chimeras containing the N carboxyl-terminal half sequence efficiently packaged N. However, the same was not true for the chimera bearing the N amino-terminal half sequence, despite its production of substantial amounts of VLPs. According to further analysis, HIV-1 NC replacement with N residues 2-213, 215-421, or 234-421 resulted in efficient VLP production at levels comparable to that of wt HIV-1, but replacement with residues 215-359, 302-421, 2-168, or 2-86 failed to restore VLP production to wild-type levels. The results suggest that the domain conferring the ability to direct VLP assembly and release in SARS-CoV N is largely contained between residues 168 and 421.
Wang
Shui-Mei
SM
Institute of Public Health, National Yang-Ming University, Taipei, Taiwan, ROC.
Chang
Yu-Fen
YF
Chen
Yi-Ming Arthur
YM
Wang
Chin-Tien
CT
D002460:Cell Line; D002499:Centrifugation, Density Gradient; D000086462:Coronavirus Nucleocapsid Proteins; D006678:HIV / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D010449:Peptide Mapping; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D013395:Sucrose / Q000737:chemistry; D014771:Virion / Q000378:metabolism*
D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; D013395:Sucrose