Strain-specific determinants of beet curly top geminivirus DNA replication

The Logan and CFH strains of the geminivirus beet curly top virus (BCTV) possess cis- and trans-DNA replication factors which exhibit specificity and are not functionally interchangeable. We demonstrate that the cis-acting replication specificity element is entirely contained within a 82- to 97-bp fragment which includes most of the viral DNA origin of replication. We also demonstrate that the strain-specific trans-acting replication determinant is located within amino acid residues 3-89 of the BCTV C1 replication protein. Transient replication assays indicated that chimeric BCTV genomes containing reciprocally exchanged regions of the CFH and Logan genomes were replication competent when the cis- and trans-replication specificity elements were derived from the same strain. Two reciprocal chimeric viral genomes with heterologous cis- and trans-replication elements were incapable of self-replication, yet could trans-replicate one another in a coinoculation experiment. Only chimeric genomes possessing the Logan trans-replication element were capable of mobilizing and amplifying a transgenic Logan derived DI-DNA. DI-DNA mobilization and amplification occurred in transient replication assays even when the helper virus genome was incapable of self-replication, providing that the trans-replication element was derived from the Logan strain. These results genetically define specific regions of the BCTV C1 replication protein determining viral DNA replication origin recognition and provide clear evidence that strains of BCTV have evolved specific cis- and trans-replication factors which functionally define the Logan and CFH strains as distinct viral agents.