29 research outputs found
Manipulating cellular factors to combat viruses: A case study from the plant eukaryotic translation initiation factors eIF4
Genes conferring resistance to plant viruses fall in two categories; the dominant genes
that mostly code for proteins with a nucleotide binding site and leucine rich repeats
(NBS-LRR), and that directly or indirectly, recognize viral avirulence factors (Avr), and the
recessive genes. The latter provide a so-called recessive resistance. They represent
roughly half of the known resistance genes and are alleles of genes that play an
important role in the virus life cycle. Conversely, all cellular genes critical for the viral
infection virtually represent recessive resistance genes. Based on the well-documented
case of recessive resistance mediated by eukaryotic translation initiation factors of
the 4E/4G family, this review is intended to summarize the possible approaches to
control viruses via their host interactors. Classically, resistant crops have been developed
through introgression of natural variants of the susceptibility factor from compatible
relatives or by random mutagenesis and screening. Transgenic methods have also been
applied to engineer improved crops by overexpressing the translation factor either in its
natural form or after directed mutagenesis. More recently, innovative approaches like
silencing or genome editing have proven their great potential in model and crop plants.
The advantages and limits of these different strategies are discussed. This example
illustrates the need to identify and characterize more host factors involved in virus
multiplication and to assess their application potential in the control of viral diseases
From a movement-deficient grapevine fanleaf virus to the identification of a new viral determinant of nematode transmission
Grapevine fanleaf virus (GFLV) and arabis mosaic virus (ArMV) are nepoviruses responsible
for grapevine degeneration. They are specifically transmitted from grapevine to grapevine by two
distinct ectoparasitic dagger nematodes of the genus Xiphinema. GFLV and ArMV move from cell to
cell as virions through tubules formed into plasmodesmata by the self-assembly of the viral movement
protein. Five surface-exposed regions in the coat protein called R1 to R5, which differ between the
two viruses, were previously defined and exchanged to test their involvement in virus transmission,
leading to the identification of region R2 as a transmission determinant. Region R4 (amino acids
258 to 264) could not be tested in transmission due to its requirement for plant systemic infection.
Here, we present a fine-tuning mutagenesis of the GFLV coat protein in and around region R4 that
restored the virus movement and allowed its evaluation in transmission. We show that residues
T258, M260, D261, and R301 play a crucial role in virus transmission, thus representing a new viral
determinant of nematode transmission
Study of the plant COPII vesicle coat subunits by functional complementation of yeast Saccharomyces cerevisiae mutants
The formation and budding of endoplasmic reticulum ER-derived vesicles depends on the COPII coat protein complex that was first identified in yeast Saccharomyces cerevisiae. The ER-associated Sec12 and the Sar1 GTPase initiate the COPII coat formation by recruiting the Sec23-Sec24 heterodimer following the subsequent recruitment of the Sec13-Sec31 heterotetramer. In yeast, there is usually one gene encoding each COPII protein and these proteins are essential for yeast viability, whereas the plant genome encodes multiple isoforms of all COPII subunits. Here, we used a systematic yeast complementation assay to assess the functionality of Arabidopsis thaliana COPII proteins. In this study, the different plant COPII subunits were expressed in their corresponding temperature-sensitive yeast mutant strain to complement their thermosensitivity and secretion phenotypes. Secretion was assessed using two different yeast cargos: the soluble alpha-factor pheromone and the membranous v-SNARE (vesicle-soluble NSF (N-ethylmaleimide-sensitive factor) attachment protein receptor) Snc1 involved in the fusion of the secretory vesicles with the plasma membrane. This complementation study allowed the identification of functional A. thaliana COPII proteins for the Sec12, Sar1, Sec24 and Sec13 subunits that could represent an active COPII complex in plant cells. Moreover, we found that AtSec12 and AtSec23 were co-immunoprecipitated with AtSar1 in total cell extract of 15 day-old seedlings of A. thaliana. This demonstrates that AtSar1, AtSec12 and AtSec23 can form a protein complex that might represent an active COPII complex in plant cells
Tubule-Guided Cell-to-Cell Movement of a Plant Virus Requires Class XI Myosin Motors
Cell-to-cell movement of plant viruses occurs via plasmodesmata (PD), organelles that evolved to facilitate intercellular communications. Viral movement proteins (MP) modify PD to allow passage of the virus particles or nucleoproteins. This passage occurs via several distinct mechanisms one of which is MP-dependent formation of the tubules that traverse PD and provide a conduit for virion translocation. The MP of tubule-forming viruses including Grapevine fanleaf virus (GFLV) recruit the plant PD receptors called Plasmodesmata Located Proteins (PDLP) to mediate tubule assembly and virus movement. Here we show that PDLP1 is transported to PD through a specific route within the secretory pathway in a myosin-dependent manner. This transport relies primarily on the class XI myosins XI-K and XI-2. Inactivation of these myosins using dominant negative inhibition results in mislocalization of PDLP and MP and suppression of GFLV movement. We also found that the proper targeting of specific markers of the Golgi apparatus, the plasma membrane, PD, lipid raft subdomains within the plasma membrane, and the tonoplast was not affected by myosin XI-K inhibition. However, the normal tonoplast dynamics required myosin XI-K activity. These results reveal a new pathway of the myosin-dependent protein trafficking to PD that is hijacked by GFLV to promote tubule-guided transport of this virus between plant cells
Structural Insights into Viral Determinants of Nematode Mediated Grapevine fanleaf virus Transmission
Many animal and plant viruses rely on vectors for their transmission from host to
host. Grapevine fanleaf virus (GFLV), a picorna-like virus from
plants, is transmitted specifically by the ectoparasitic nematode
Xiphinema index. The icosahedral capsid of GFLV, which
consists of 60 identical coat protein subunits (CP), carries the determinants of
this specificity. Here, we provide novel insight into GFLV transmission by
nematodes through a comparative structural and functional analysis of two GFLV
variants. We isolated a mutant GFLV strain (GFLV-TD) poorly transmissible by
nematodes, and showed that the transmission defect is due to a glycine to
aspartate mutation at position 297 (Gly297Asp) in the CP. We next determined the
crystal structures of the wild-type GFLV strain F13 at 3.0 Ã… and of
GFLV-TD at 2.7 Ã… resolution. The Gly297Asp mutation mapped to an exposed
loop at the outer surface of the capsid and did not affect the conformation of
the assembled capsid, nor of individual CP molecules. The loop is part of a
positively charged pocket that includes a previously identified determinant of
transmission. We propose that this pocket is a ligand-binding site with
essential function in GFLV transmission by X. index. Our data
suggest that perturbation of the electrostatic landscape of this pocket affects
the interaction of the virion with specific receptors of the nematode's
feeding apparatus, and thereby severely diminishes its transmission efficiency.
These data provide a first structural insight into the interactions between a
plant virus and a nematode vector
A Family of Plasmodesmal Proteins with Receptor-Like Properties for Plant Viral Movement Proteins
Plasmodesmata (PD) are essential but poorly understood structures in plant cell walls that provide symplastic continuity and intercellular communication pathways between adjacent cells and thus play fundamental roles in development and pathogenesis. Viruses encode movement proteins (MPs) that modify these tightly regulated pores to facilitate their spread from cell to cell. The most striking of these modifications is observed for groups of viruses whose MPs form tubules that assemble in PDs and through which virions are transported to neighbouring cells. The nature of the molecular interactions between viral MPs and PD components and their role in viral movement has remained essentially unknown. Here, we show that the family of PD-located proteins (PDLPs) promotes the movement of viruses that use tubule-guided movement by interacting redundantly with tubule-forming MPs within PDs. Genetic disruption of this interaction leads to reduced tubule formation, delayed infection and attenuated symptoms. Our results implicate PDLPs as PD proteins with receptor-like properties involved the assembly of viral MPs into tubules to promote viral movement
The specific transmission of Grapevine fanleaf virus by its nematode vector Xiphinema index is solely determined by the viral coat protein
AbstractThe viral determinants involved in the specific transmission of Grapevine fanleaf virus (GFLV) by its nematode vector Xiphinema index are located within the 513 C-terminal residues of the RNA2-encoded polyprotein, that is, the 9 C-terminal amino acids of the movement protein (2BMP) and contiguous 504 amino acids of the coat protein (2CCP) [Virology 291 (2001) 161]. To further delineate the viral determinants responsible for the specific spread, the four amino acids that are different within the 9 C-terminal 2BMP residues between GFLV and Arabis mosaic virus (ArMV), another nepovirus which is transmitted by Xiphinema diversicaudatum but not by X. index, were subjected to mutational analysis. Of the recombinant viruses derived from transcripts of GFLV RNA1 and RNA2 mutants that systemically infected herbaceous host plants, all with the 2CCP of GFLV were transmitted by X. index unlike none with the 2CCP of ArMV, regardless of the mutations within the 2BMP C-terminus. These results demonstrate that the coat protein is the sole viral determinant for the specific spread of GFLV by X. index
Symptom Determinants of Grapevine Fanleaf Virus in Nicotiana Species
International audienc
The 50 distal amino acids of the 2A HP homing protein of Grapevine fanleaf virus elicit a hypersensitive reaction on Nicotiana occidentalis
International audienceAvirulence factors are critical for the arm's race between a virus and its host in determining incompatible reactions. The response of plants to viruses from the genus Nepovirus in the family Secoviridae, including Grapevine fanleaf virus (GFLV), is well characterized, although the nature and characteristics of the viral avirulence factor remain elusive. By using infectious clones of GFLV strains F13 and GHu in a reverse genetics approach with wild-type, assortant and chimeric viruses, the determinant of necrotic lesions caused by GFLV-F13 on inoculated leaves of Nicotiana occidentalis was mapped to the RNA2-encoded protein 2A(HP), particularly to its 50 C-terminal amino acids. The necrotic response showed hallmark characteristics of a genuine hypersensitive reaction, such as the accumulation of phytoalexins, reactive oxygen species, pathogenesis-related protein 1c and hypersensitivity-related (hsr) 203J transcripts. Transient expression of the GFLV-F13 protein 2A(HP) fused to an enhanced green fluorescent protein (EGFP) tag in N.occidentalis by agroinfiltration was sufficient to elicit a hypersensitive reaction. In addition, the GFLV-F13 avirulence factor, when introduced in GFLV-GHu, which causes a compatible reaction on N.occidentalis, elicited necrosis and partially restricted the virus. This is the first identification of a nepovirus avirulence factor that is responsible for a hypersensitive reaction in both the context of virus infection and transient expression