NTP binding induces conformational changes in the double-stranded RNA bacteriophage ø6 subviral particles.

Bacteriophage ø6 is a double-stranded RNA virus consisting of a nucleocapsid (NC) surrounded by a membrane. Beneath the NC major coat protein, P8, resides the ø6 RNA polymerase complex which is composed of four early proteins P1, P2, P4, and P7. Protein P1 forms the dodecahedral framework with which the other three proteins are associated. We have developed a new method for the isolation of stable polymerase complex particles which retain their structural integrity and polymerase activity for several days. Purine nucleotides, especially GTP, dGTP, ddGTP, and GDP, stabilized the particle efficiently. Furthermore, binding of any NTP was shown to induce conformational changes in the NC structure, as detected by alterations in the binding properties of NC-specific monoclonal antibodies. In the presence of NTPs, most of the epitopes in protein P4 become more exposed than without NTPs, while the epitopes in protein P8 were either masked or unmasked due to NTP binding. Based on the accessibility of the epitopes of protein P1 on the NC, we postulate that at least part of this protein is also accessible on the NC surface.