TY - JOUR
T1 - The intracellular sites of early replication and budding of SARS-coronavirus
AU - Stertz, Silke
AU - Reichelt, Mike
AU - Spiegel, Martin
AU - Kuri, Thomas
AU - Martínez-Sobrido, Luis
AU - García-Sastre, Adolfo
AU - Weber, Friedemann
AU - Kochs, Georg
N1 - Funding Information:
We thank S. Becker for providing the FFM-1 strain of SARS-CoV, S. Baker, H-P. Hauri, J. Simpson and T. Yoshimori for antibodies and plasmids, J. Paragas for SARS-CoV RNA and J. Krijnse-Locker, G. Griffith (EMBL, Heidelberg) and John Perrino (Stanford, CSIF) for training and advice concerning electron microscopy and K. Kirkegaard for supporting this collaboration and for helpful discussion. Furthermore, we want to acknowledge B. Moss for providing us with the pRB21 plasmid as well as the vRB21 vaccinia strain to generate recombinant vaccinia virus, as well as A. Pritsker at Mount Sinai Hybridoma Center for generating the monoclonal antibody and R. Cádagan for excellent technical assistance. We thank O. Haller and P. Staeheli for helpful discussions and critical comments on the manuscript. This work was supported by grants from the Sino-German Center for Research Promotion (FW) and the Deutsche Forschungsgemeinschaft (FW) as well as by NIH grants (AG-S) and the Dean's fellowship from Stanford University (MR).
PY - 2007/5/10
Y1 - 2007/5/10
N2 - In this study, we analyzed the replication and budding sites of severe acute respiratory syndrome coronavirus (SARS-CoV) at early time points of infection. We detected cytoplasmic accumulations containing the viral nucleocapsid protein, viral RNA and the non-structural protein nsp3. Using EM techniques, we found that these putative viral replication sites were associated with characteristic membrane tubules and double membrane vesicles that most probably originated from ER cisternae. In addition to its presence at the replication sites, N also accumulated in the Golgi region and colocalized with the viral spike protein. Immuno-EM revealed that budding occurred at membranes of the ERGIC (ER-Golgi intermediate compartment) and the Golgi region as early as 3 h post infection, demonstrating that SARS-CoV replicates surprisingly fast. Our data suggest that SARS-CoV establishes replication complexes at ER-derived membranes. Later on, viral nucleocapsids have to be transported to the budding sites in the Golgi region where the viral glycoproteins accumulate and particle formation occurs.
AB - In this study, we analyzed the replication and budding sites of severe acute respiratory syndrome coronavirus (SARS-CoV) at early time points of infection. We detected cytoplasmic accumulations containing the viral nucleocapsid protein, viral RNA and the non-structural protein nsp3. Using EM techniques, we found that these putative viral replication sites were associated with characteristic membrane tubules and double membrane vesicles that most probably originated from ER cisternae. In addition to its presence at the replication sites, N also accumulated in the Golgi region and colocalized with the viral spike protein. Immuno-EM revealed that budding occurred at membranes of the ERGIC (ER-Golgi intermediate compartment) and the Golgi region as early as 3 h post infection, demonstrating that SARS-CoV replicates surprisingly fast. Our data suggest that SARS-CoV establishes replication complexes at ER-derived membranes. Later on, viral nucleocapsids have to be transported to the budding sites in the Golgi region where the viral glycoproteins accumulate and particle formation occurs.
KW - Budding
KW - Double membrane vesicles
KW - Replication
KW - SARS-coronavirus
UR - http://www.scopus.com/inward/record.url?scp=34147135956&partnerID=8YFLogxK
U2 - 10.1016/j.virol.2006.11.027
DO - 10.1016/j.virol.2006.11.027
M3 - Article
C2 - 17210170
AN - SCOPUS:34147135956
SN - 0042-6822
VL - 361
SP - 304
EP - 315
JO - Virology
JF - Virology
IS - 2
ER -