GAPDH-A Recruits a Plant Virus Movement Protein to Cortical Virus Replication Complexes to Facilitate Viral Cell-to-Cell Movement

• DOI: 10.1371/journal.ppat.1004505
• Publication Date: 2014-11-20T19:21:10Z
• Authors: Masanori Kaido; Kazutomo Abe; Akira Mine; Kiwamu Hyodo; Takako Taniguchi; Hisaaki Taniguchi; Kazuyuki Mise; Tetsuro Okuno
• Keywords: Movement protein; Bimolecular fluorescence complementation; Tandem affinity purification; Protein-fragment complementation assay

The formation of virus movement protein (MP)-containing punctate structures on the cortical endoplasmic reticulum is required for efficient intercellular Red clover necrotic mosaic (RCNMV), a bipartite positive-strand RNA plant virus. We found that these constitute viral replication complex (VRC) in addition to previously reported aggregate formed adjacent nucleus. identified host proteins interacted with RCNMV MP virus-infected Nicotiana benthamiana leaves using tandem affinity purification method followed by mass spectrometry. One was glyceraldehyde 3-phosphate dehydrogenase-A (NbGAPDH-A), which component Calvin-Benson cycle chloroplasts. Virus-induced gene silencing NbGAPDH-A reduced multiplication inoculated leaves, but not single cells, thereby suggesting GAPDH-A plays positive role cell-to-cell RCNMV. fusion and green fluorescent localized exclusively In presence RNA1, however, VRC as well Bimolecular fluorescence complementation assay GST pulldown confirmed vivo vitro interactions, respectively, between NbGAPDH-A. Furthermore, inhibited localization VRC. discuss possible roles process.