Mutagenesis of the BC1 and BV1 genes of African cassava mosaic virus identifies conserved amino acids that are essential for spread

The products of three open reading frames encoded by the bipartite geminiviruses have been implicated in viral spread: AC2, BV1 and BC1. Alignment of the DNA B encoded gene products, BV1 and BC1, from African cassava mosaic virus (ACMV) with six other bipartite geminiviruses showed several highly conserved regions. Specific amino acids were selected for mutagenic studies to ascertain the tolerance of the virus to change and to identify the regions within these two proteins required for normal functioning. Various mutant DNA B constructs, and a wild-type construct, were inoculated onto three host plant species with an equivalent DNA A construct. Three of the mutant constructs were infectious on Nicotiana benthamiana and N. clevelandii, but only two induced ACMV disease symptoms on N. tabacum cv. Samsun. Sequencing of the viral DNA extracted from the sap of systemically infected plants confirmed the maintenance of introduced base changes. The amino acid at position 95 on the BV1 gene product was identified as non-essential for normal functioning of the protein. The alteration of the amino acid at position 145 in BC1 demonstrated the ability of the virus to tolerate a conservative change. The lack of tolerance to other changes in amino acids has given an indication of the importance of maintaining protein structure for these proteins to function normally.