Abstract
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 132168) and the highly acidic C-terminal region (aa 236241) of the P protein were found to be essential for N-P interaction. In addition, the highly basic N-terminal region (amino acids 140) was found to be involved in N-P interaction to a lesser extent.
* Author for correspondence. Fax +1 301 935 6079. e-mail SS5@umail.umd.edu
† Present address: Laboratory of Clinical Investigation, NIAID, National Institutes of Health, Bethesda, MD 20892, USA.
‡ Present address: Insect Neurobiology and Hormone Laboratory, USDA, ARS, Beltsville, MD 20705, USA.