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Identification of protein interactions of grapevine fanleaf virus RNA-dependent RNA polymerase during infection of Nicotiana benthamiana by affinity purification and tandem mass spectrometry.

ABSTRACT: The RNA-dependent RNA polymerase (1E^Pol) is involved in replication of grapevine fanleaf virus (GFLV, Nepovirus, Secoviridae) and causes vein clearing symptoms in Nicotiana benthamiana. Information on protein 1E^Pol interaction with other viral and host proteins is scarce. To study protein 1E^Pol biology, three GFLV infectious clones, i.e. GHu (a symptomatic wild-type strain), GHu-1E_K802G (an asymptomatic GHu mutant) and F13 (an asymptomatic wild-type strain), were engineered with protein 1E^Pol fused to a V5 epitope tag at the C-terminus. Following Agrobacterium tumefaciens-mediated delivery of GFLV clones in N. benthamiana and protein extraction at seven dpi, when optimal 1E^Pol:V5 accumulation was detected, two viral and six plant putative interaction partners of V5-tagged protein 1E^Pol were identified for the three GFLV clones by affinity purification and tandem mass spectrometry. This study provides insights into the protein interactome of 1E^Pol during GFLV systemic infection in N. benthamiana and lays the foundation for validation work.

SUBMITTER: Osterbaan LJ

PROVIDER: S-EPMC8295916 | biostudies-literature |

REPOSITORIES: biostudies-literature

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: Proteomic analysis of Nicotiana benthamiana upon infection of grapevine fanleaf virus strains

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