Analysis of severe acute respiratory syndrome coronavirus structural proteins in virus-like particle assembly.
SARS-CoV has four major structural proteins: the N, S, M, and E proteins. To investigate the mechanism of SARS-CoV assembly, we cloned the genes encoding these four proteins into the eukaryotic expression vector pCAGGS and transfected them into 293T cells. When all four expression vectors were co-transfected VLP formed, as confirmed using electron microscopy. Using a rabbit polyclonal antibody specific to the N protein, N-protein-containing particles similar in size to the VLP were also observed by immunoelectron microscopy, indicating that the VLP contained the N protein. Co-immunoprecipitation analyses demonstrated an interaction between the N and M proteins, suggesting that N protein binds directly to M protein to be incorporated into VLP.
Nakauchi
Mina
M
Laboratory of Public Health, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido, Japan.
Kariwa
Hiroaki
H
Kon
Yasuhiro
Y
Yoshii
Kentaro
K
Maeda
Akihiko
A
Takashima
Ikuo
I
D002460:Cell Line; D006801:Humans; D011485:Protein Binding; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D045169:Severe Acute Respiratory Syndrome / Q000821:virology*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism*; D019065:Virus Assembly
D015678:Viral Structural Proteins