Structures of fd Gene 5 Protein·Nucleic Acid Complexes: A Combined Solution Scattering and Electron Microscopy Study

Abstract

Small-angle scattering and electron microscopy studies of fd gene 5 protein (g5p) and reconstituted g5p·nucleic acid complexes have been used to test models for the complexes and evaluate their uniqueness. In addition, we have obtained new information on the dependence of nucleotide type and protein/nucleotide (P/N) ratio on the structure of the complexes. Reconstituted complexes were made with single-stranded fd viral DNA (fd ssDNA), poly[d(A)] and poly[r(A)]. All complexes form similar left-handed, flexible superhelices having approximately the same diameter, but the pitch differs among these complexes. The g5p protein is a dimer in solution and the dimers associate to form a superhelical framework to which the polynucleotide is attached. The combined X-ray and neutron scattering data confirm the nucleic acid is inside the protein superhelix. A Monte Carlo integration modeling procedure applied to the scattering data was used to systematically test large numbers of possible models for each complex, and previously proposed models based on parameters obtained from electron microscopy were found to be essentially correct and unique. The data on the complexes with differentP/νratios showed that mass per unit length values decreased while the rise per dimer and pitch of the superhelix increased for g5p·fd-ssDNA complexes with decreasingP/Nratios.