Mapping the domains on the phosphoprotein of bovine respiratory syncytial virus required for N-P interaction using a two-hybrid system.
Specific interactions between the nucleocapsid protein (N) and the phosphoprotein (P) of bovine respiratory syncytial virus (BRSV) have been investigated using a yeast-based two-hybrid system. Plasmids encoding the yeast GAL4 DNA binding domain fused with the N gene and GAL4 activation domain fused with the P gene were cotransfected into competent yeast cells. The ability of the N and P proteins to interact in vivo was measured by activation of the lacZ reporter gene by the GAL4 transactivation region. Results indicated that the N and P proteins interact very strongly in vivo. When interactions between N and various deletion mutants of the P protein were examined, an internal region (aa 132-168) and the highly acidic C-terminal region (aa 236-241) of the P protein were found to be essential for N-P interaction. In addition, the highly basic N-terminal region (amino acids 1-40) was found to be involved in N-P interaction to a lesser extent.
Mallipeddi
S K
SK
VA-MD Regional College of Veterinary Medicine, University of Maryland, College Park 20742, USA.
Lupiani
B
B
Samal
S K
SK
D001665:Binding Sites; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D010750:Phosphoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D018112:Respiratory Syncytial Virus, Bovine / Q000737:chemistry*; D013329:Structure-Activity Relationship; D014162:Transfection; D014758:Viral Core Proteins / Q000737:chemistry / Q000378:metabolism*
D010750:Phosphoproteins; D014758:Viral Core Proteins