To identify host factors that play critical roles in processes, including cell-to-cell movement of plant-adapted rhabdovi- ruses, we constructed and validated a high-resolution Nico- tiana benthamiana yeast two-hybrid library. The library was screened with the putative movement protein (sc 4), nu- cleocapsid (N), and matrix (M) proteins of Sonchus yellow net virus (SYNV). This resulted in identification of 31 potential host factors. Steady-state localization studies using autofluorescent protein fusions to full-length clones of inter- actors were conducted in transgenic N. benthamiana marker lines. Bimolecular fluorescence complementation assays were used to validate two-hybrid interactions. The sc4 inter- actor, sc4i21, localized to microtubules. The N interactor, Ni67, localized to punctuate loci on the endoplasmic reticulum. These two proteins are 84% identical homologues of the Arabidopsis phloem-associated transcription activator AtVOZl, and contain functional nuclear localization signals. Sc4i17 is a microtubule-associated motor protein. The M interactor, Mi7, is a nuclear-localized transcription factor. Combined with a binary interaction map for SYNV proteins, our data support a model in which the SYNV nu- cleocapsids are exported from the nucleus and moved cell- to-cell by transcription activators tethered in the cytoplasm.
|Number of pages||13|
|Journal||Molecular Plant-Microbe Interactions|
|State||Published - Nov 2010|
ASJC Scopus subject areas
- Agronomy and Crop Science