Start Date: Mar 15, 2012
End Date: Mar 14, 2015
We have shown the CP without RTP can assemble virions, but they are movement impaired in both plants and aphids. CP and RTP act together to regulate virus movement and survival in the aphid. RTP also functions as a soluble protein to confine virus to phloem tissues. P17, a nonstructural virus movement protein, acts in a host dependent manner and affects virus uptake during aphid feeding. Recently, we have identified a number of aphid and aphid symbiont proteins that are linked to virus transmission. Also we have identified a number of plant proteins that co-purify with transmissible virus, but are absent from virus that cannot move through the aphid. The logical extension of this work is to identify which of the aphid and plant candidate proteins are truly interacting with which virus protein and to determine how these various proteins orchestrate virus movement in the plant and aphid to ensure maximum transmission and continued virus existence. Our approach will focus on the use of the latest techniques in targeted proteomics and cell biology to identify aphid, symbiont and plant proteins that interact with each of the virus proteins. Mutations in the virus and plant proteins that disrupt these interactions will be used to identify how and where each protein functions independent of, and in concert with, other virus proteins in both plants and aphids. In vitro feeding and microinjection techniques allow us to mix and match various virions and proteins for study in various aphid tissues. GFP tagged proteins will be used to localize interactions to specific tissues and cell organelles.