Effect of a single amino acid substitution in the NLS domain of Tomato yellow leaf curl virus-Israel (TYLCV-IL) capsid protein (CP) on its activity and on the virus life cycle

The capsid protein (CP) of Tomato yellow leaf curl virus-Israel (TYLCV-IL), encoded by the v1 gene, is the only known component of the viral capsid. Three point mutations introduced into the conserved NLS region of the CP were investigated. One mutant, in which the Arg at position 19 was converted to Leu, had the most significant effect on the CP-CP homotypic interaction as well as on CP's interaction with its nuclear receptor karyopherin α1 and with the protein GroEL. The latter has been suggested to protect the virions in the insect vector hemolymph. These effects were first observed by yeast two-hybrid assay and then confirmed in tobacco protoplasts by measuring fluorescence resonance energy transfer (FRET) between YFP- and CFP-tagged proteins. Most importantly, when the point mutation converting Arg 19 to Leu was introduced into the full-length TYLCV genome, it disrupted its ability to cause symptoms.