Characterization of protein-protein interactions between the nucleocapsid protein and membrane protein of the SARS coronavirus.
The human coronavirus, associated with severe acute respiratory syndrome (SARS-CoV), was identified and molecularly characterized in 2003. Sequence analysis of the virus indicates that there is only 20% amino acid (aa) identity with known coronaviruses. Previous studies indicate that protein-protein interactions amongst various coronavirus proteins are critical for viral assembly. Yet, little sequence homology between the newly identified SARS-CoV and those previously studied coronaviruses suggests that determination of protein-protein interaction and identification of amino acid sequences, responsible for such interaction in SARS-CoV, are necessary for the elucidation of the molecular mechanism of SARS-CoV replication and rationalization of anti-SARS therapeutic intervention. In this study, we employed mammalian two-hybrid system to investigate possible interactions between SARS-CoV nucleocapsid (N) and the membrane (M) proteins. We found that interaction of the N and M proteins takes place in vivo and identified that a stretch of amino acids (168-208) in the N protein may be critical for such protein-protein interactions. Importantly, the same region has been found to be required for multimerization of the N protein (He et al., 2004) suggesting this region may be crucial in maintaining correct conformation of the N protein for self-interaction and interaction with the M protein.
He
Runtao
R
National Microbiology Laboratory, Health Canada, 1015 Arlington St., Winnipeg, Man., Canada R3E 3R2. runtao_he@hc-sc.gc.ca
Leeson
Andrew
A
Ballantine
Melissa
M
Andonov
Anton
A
Baker
Lindsay
L
Dobie
Frederick
F
Li
Yan
Y
Bastien
Nathalie
N
Feldmann
Heinz
H
Strocher
Ute
U
Theriault
Steven
S
Cutts
Todd
T
Cao
Jingxin
J
Booth
Timothy F
TF
Plummer
Frank A
FA
Tyler
Shaun
S
Li
Xuguang
X
D001665:Binding Sites; D000086862:Coronavirus M Proteins; D000086462:Coronavirus Nucleocapsid Proteins; D019590:Nucleocapsid Proteins / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D045473:Severe acute respiratory syndrome-related coronavirus / Q000378:metabolism* / Q000502:physiology; D017384:Sequence Deletion; D020798:Two-Hybrid System Techniques; D014763:Viral Matrix Proteins / Q000378:metabolism*; D014764:Viral Proteins / Q000378:metabolism; D019065:Virus Assembly
D000086862:Coronavirus M Proteins; D000086462:Coronavirus Nucleocapsid Proteins; C000706237:M protein, SARS-CoV; D019590:Nucleocapsid Proteins; D014763:Viral Matrix Proteins; D014764:Viral Proteins