We will investigate the attachment of the genome of bacteriophage 029 at various regions of the Bacillus subtilis cell membrane. These include membrane co-sedimenting with two forms of folded bacterial chromosomes (nucleoids), membrane regions derived from the mesosomes, and membrane at the poles of the cell obtained by isolating minicells from division mutants. The protein composition in DNA - membrane complexes will be characterized and compared which contain host genome: membrane attachment sites, and viral genome: membrane attachment sites. In viral DNA: membrane complexes, the interaction of the viral genome and viral proteins with membrane proteins will be determined by chemical cross-linking using lipid soluble and water soluble bifunctional agents, and through photochemically induced cross-linking of viral DNA substituted with brom deoxyuridine. We hope that the experiments outlined in this proposal will describe the molecular composition of viral and host genome attachment site at the cell membrane and allow us to define the role of the viral specific protein (the product of gene 2) required for DNA replication and membrane attachment.
NIH ExPorter
NIH ExPorter