Abstract
The eukaryotic translation elongation factor 1 (eEF1) has two components: the G-protein eEF1A and the nucleotide exchange factor eEF1B. In plants, eEF1B is itself composed of a structural protein (eEF1Bγ) and two nucleotide exchange subunits (eEF1Bα and eEF1Bβ). To test the effects of elongation factors on virus infection, we isolated eEF1A and eEF1B genes from pepper (Capsicum annuum) and suppressed their homologs in Nicotiana benthamiana using virus-induced gene silencing (VIGS). The accumulation of a green fluorescent protein (GFP)-tagged Potato virus X (PVX) was significantly reduced in the eEF1Bβ- or eEF1Bɣ-silenced plants as well as in eEF1A-silenced plants. Yeast two-hybrid and co-immunoprecipitation analyses revealed that eEF1Bα and eEF1Bβ interacted with eEF1A and that eEF1A and eEF1Bβ interacted with triple gene block protein 1 (TGBp1) of PVX. These results suggest that both eEF1A and eEF1Bβ play essential roles in the multiplication of PVX by physically interacting with TGBp1. Furthermore, using eEF1Bβ deletion constructs, we found that both N- (1-64 amino acids) and C-terminal (150-195 amino acids) domains of eEF1Bβ are important for the interaction with PVX TGBp1 and that the C-terminal domain of eEF1Bβ is involved in the interaction with eEF1A. These results suggest that eEF1Bβ could be a potential target for engineering virus-resistant plants.
Highlights
Plant viruses are excellent subjects for the elucidation of host—microbe interactions because they are simple and obligate intracellular parasites and are entirely dependent on host cellular metabolism for their life cycle
Using eEF1Bβ deletion constructs, we found that both N- (1-64 amino acids) and C-terminal (150-195 amino acids) domains of eEF1Bβ are important for the interaction with Potato virus X (PVX) triple gene block protein 1 (TGBp1) and that the C-terminal domain of eEF1Bβ is involved in the interaction with eEF1A
Using yeast two-hybrid and co-immunoprecipitation analyses, we demonstrated that eEF1A interacts with eEF1Bα and eEF1Bβ, and that each eEF1A and eEF1Bβ interacts with the TGBp1 protein of PVX
Summary
Plant viruses are excellent subjects for the elucidation of host—microbe interactions because they are simple and obligate intracellular parasites and are entirely dependent on host cellular metabolism for their life cycle. Roles of Plant eEF1B Subunits in PVX Infection. California, Davis) for the TRV-LIC vector, K.H. Kim (Seoul National University) for PVX-GFP and D. Choi (Seoul National University) for access to the C. annuum sequence database
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