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

Masanori Kaido, Kazutomo Abe, Akira Mine, Kiwamu Hyoudou, Takako Taniguchi, Hisaaki Taniguchi, Kazuyuki Mise, Tetsuro Okuno

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

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

Original languageEnglish
JournalPLoS Pathogens
Volume10
Issue number11
DOIs
Publication statusPublished - Nov 1 2014
Externally publishedYes

Fingerprint

Plant Viral Movement Proteins
Tombusviridae
Virus Replication
Cell Movement
Proteins
Chloroplasts
Gene Silencing
Viruses
Plant Viruses
Glyceraldehyde-3-Phosphate Dehydrogenases
RNA Viruses
Photosynthesis
Green Fluorescent Proteins
Endoplasmic Reticulum
Tobacco
Mass Spectrometry
Fluorescence

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology
  • Medicine(all)

Cite this

GAPDH-A Recruits a Plant Virus Movement Protein to Cortical Virus Replication Complexes to Facilitate Viral Cell-to-Cell Movement. / Kaido, Masanori; Abe, Kazutomo; Mine, Akira; Hyoudou, Kiwamu; Taniguchi, Takako; Taniguchi, Hisaaki; Mise, Kazuyuki; Okuno, Tetsuro.

In: PLoS Pathogens, Vol. 10, No. 11, 01.11.2014.

Research output: Contribution to journalArticle

Kaido, Masanori ; Abe, Kazutomo ; Mine, Akira ; Hyoudou, Kiwamu ; Taniguchi, Takako ; Taniguchi, Hisaaki ; Mise, Kazuyuki ; Okuno, Tetsuro. / GAPDH-A Recruits a Plant Virus Movement Protein to Cortical Virus Replication Complexes to Facilitate Viral Cell-to-Cell Movement. In: PLoS Pathogens. 2014 ; Vol. 10, No. 11.
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