1887

Abstract

We have devised an system for functional analysis of the replication-associated protein (Rep) of African cassava mosaic virus (ACMV). Using this assay and PCR-based random mutagenesis, we have identified an ACMV Rep mutant that failed to trigger the hypersensitive response (HR), but had an enhanced ability to initiate DNA replication. The mutant Rep–green fluorescent protein (GFP) fusion protein was localized to the nucleus. Sequence analysis showed that the mutated gene had three nucleotide changes (A6→T, T375→G and G852→A); only the A6→T transversion resulted in an amino acid substitution (Arg to Ser), which is at the second residue in the 358 amino acid ACMV Rep protein. Our results indicate that a single amino acid can alter the differential ability of ACMV Rep to trigger the host-mediated HR defence mechanism and to initiate viral DNA replication. The implications of this finding are discussed in the context of plant–virus interactions.

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2008-10-01
2020-01-24
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