Binding analyses for the interaction between plant virus genome-linked protein (VPg) and plant translational initiation factors

Mizumoto, Hiroshi; Suehiro, Noriko; Tomoo, Koji; Muto, Shinji; Takahashi, Tsubasa; Tsukamoto, Tomoko; Ohmori, T.; Natsuaki, Tomohide

doi:10.1016/j.biochi.2005.09.002

Cited by 73 publications

(68 citation statements)

References 70 publications

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“…It has been noted that the eIF4E proteins encoded by the pvr1 allele from pepper and the sbm1 allele from pea, which both contain the G107R substitution, show reduced cap binding (Gao et al, 2004;Kang et al, 2005a). Recent studies have suggested that the mRNA cap structure and potyvirus VPg at least partially share the protein binding pocket structure of the eIF4E protein or its isoform eIF(iso)4E (Michon et al, 2006;Miyoshi et al, 2006).…”

Section: The Vpg Binding and Cap Binding Abilities Of Eif4e Are Strucmentioning

confidence: 99%

“…The physical interaction between host eukaryotic initiation factor eIF4E or eIF(iso)4E and the viral genome-linked protein (VPg) is critical for viral infection by several members of the genus potyvirus (Wittmann et al, 1997;Leonard et al, 2000;Grzela et al, 2006;Miyoshi et al, 2006;Robaglia and Caranta, 2006). The major role of eIF4E in the host cell is initiating protein translation by allowing recognition and interaction with the cap structure of cellular mRNA.…”

Section: Introductionmentioning

confidence: 99%

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Functional Dissection of Naturally Occurring Amino Acid Substitutions in eIF4E That Confers Recessive Potyvirus Resistance in Plants

et al. 2007

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Naturally existing variation in the eukaryotic translation initiation factor 4E (eIF4E) homolog encoded at the pvr1 locus in Capsicum results in recessively inherited resistance against several potyviruses. Previously reported data indicate that the physical interaction between Capsicum-eIF4E and the viral genome-linked protein (VPg) is required for the viral infection in the Capsicum-Tobacco etch virus (TEV) pathosystem. In this study, the potential structural role(s) of natural variation in the eIF4E protein encoded by recessive resistance alleles and their biological consequences have been assessed. Using highresolution three-dimensional structural models based on the available crystallographic structures of eIF4E, we show that the amino acid substitution G107R, found in many recessive plant virus resistance genes encoding eIF4E, is predicted to result in a substantial modification in the protein binding pocket. The G107R change was shown to not only be responsible for the interruption of VPg binding in planta but also for the loss of cap binding ability in vitro, the principal function of eIF4E in the host. Overexpression of the Capsicum-eIF4E protein containing the G107R amino acid substitution in Solanum lycopersicum indicated that this polymorphism alone is sufficient for the acquisition of resistance against several TEV strains.

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Section: The Vpg Binding and Cap Binding Abilities Of Eif4e Are Strucmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Functional Dissection of Naturally Occurring Amino Acid Substitutions in eIF4E That Confers Recessive Potyvirus Resistance in Plants

et al. 2007

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“…Similar interactions are taking place between the VPg of other potyviruses and either eIF(iso) 4E or eIF4E (20,46). The cap analogue m 7 GDP and VPg bound to eIF4E or eIF(iso)4E at two distinct sites, although binding of one ligand to the translation factor reduced its affinity for the other ligand (31,33,34). This suggests that the potyviral VPg interferes with the formation of a translational initiation complex on cellular mRNAs by sequestering the translation factor.…”

mentioning

confidence: 93%

Visualization of the Interaction between the Precursors of VPg, the Viral Protein Linked to the Genome ofTurnip Mosaic Virus, and the Translation Eukaryotic Initiation Factor iso 4E In Planta

Beauchemin

Boutet

Laliberté

2007

J Virol

150

116

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The RNA genome of Turnip mosaic virus is covalently linked at its 5 end to a viral protein known as VPg. This protein binds to the translation eukaryotic initiation factor iso 4E [eIF(iso)4E]. This interaction has been shown to be important for virus infection, although its exact biological function(s) has not been elucidated. In this study, we investigated the subcellular site of the VPg-eIF(iso)4E interaction using bimolecular fluorescence complementation (BiFC). As a first step, eIF(iso)4E, 6K-VPg-Pro, and VPg-Pro were expressed as full-length green fluorescent protein (GFP) fusions in Nicotiana benthamiana, and their subcellular localizations were visualized by confocal microscopy. eIF(iso)4E was predominantly associated with the endoplasmic reticulum (ER), and VPg-Pro was observed in the nucleus and possibly the nucleolus, while 6K-VPg-Pro-GFP induced the formation of cytoplasmic vesicles budding from the ER. In BiFC experiments, reconstituted green fluorescence was observed throughout the nucleus, with a preferential accumulation in subnuclear structures when the GFP split fragments were fused to VPg-Pro and eIF(iso)4E. On the other hand, the interaction of 6K-VPg-Pro with eIF(iso)4E was observed in cytoplasmic vesicles embedded in the ER. These data suggest that the association of VPg with the translation factor might be needed for two different functions, depending of the VPg precursor involved in the interaction. VPg-Pro interaction with eIF(iso)4E may be involved in perturbing normal cellular functions, while 6K-VPg-Pro interaction with the translation factor may be needed for viral RNA translation and/or replication.One of the initial steps in protein synthesis is the recruitment of mRNAs by the translation eukaryotic initiation factor 4F (eIF4F) complex (17). In plants, this complex is composed of two proteins, eIF4E and eIF4G (4). eIF4E is the cap-binding protein, while eIF4G is a large polypeptide containing binding sites for eIF4E, eIF4A, eIF3, and the poly(A) binding protein. eIF4F simultaneously interacts with the cap structure of mRNA (through eIF4E) and the ribosome-associated eIF3 (through eIF4G). Thus, eIF4F executes the pivotal function of bridging mRNAs with ribosomes. Another cap-binding complex, eIF(iso)4F, has been identified in plants (5) and is composed of a smaller eIF(iso)4E subunit and a larger eIF(iso)4G subunit relative to the eIF4F subunits. Those two complexes perform essentially the same task in translation but have different affinities for certain classes of mRNA substrates and may be involved in different cellular events (14).Potyviruses have a positive-strand RNA genome of approximately 10 kb (53). The genome codes for a single long polyprotein that is processed by viral proteinases into 10 mature proteins. The viral RNA is polyadenylated at its 3Ј end and has a covalently linked virus-encoded protein (VPg) instead of a cap structure at the 5Ј end. Being derived from a polyprotein, VPg exists in various forms (i.e., VPg, VPg-Pro, 6K-VPg-Pro) that fulfill specific functions....

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“…Moreover, it has been shown that VPgs can directly regulate the protease activity as SeMV protease is active in trans only in fusion with VPg (Satheshkumar et al, 2005). To perform their various functions, VPgs establish interactions with several viral or host partners such as VPg itself, nuclear inclusion protein b, helper component protease, cylindrical inclusion protein, coat protein or eukaryotic translation initiation factors: eIF4E, eIF4G, eIF4A, eIF3 and the poly(A)-binding protein (Daughenbaugh et al, 2003(Daughenbaugh et al, , 2006Goodfellow et al, 2005;Hébrard et al, 2010;Khan et al, 2008;Lin et al, 2009;Michon et al, 2006;Miyoshi et al, 2006). For RYMV, an interaction of VPg with eIF(iso)4G is known to be crucial for virus infection (Albar et al, 2006;Hébrard et al, 2006Hébrard et al, , 2010.…”

Section: Introductionmentioning

confidence: 99%

Protein-RNA linkage and post-translational modifications of two sobemovirus VPgs

Olspert

Peil

Hébrard

et al. 2010

Journal of General Virology

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Sobemoviruses possess a viral genome-linked protein (VPg) attached to the 59 end of viral RNA. VPg is processed from the viral polyprotein. In the current study, Cocksfoot mottle virus (CfMV) and Rice yellow mottle virus (RYMV) VPgs were purified from virions and analysed by mass spectrometry. The cleavage sites in the polyprotein and thereof the termini of VPg were experimentally proven. The lengths of the mature VPgs were determined to be 78 and 79 aa residues, respectively. The amino acid residues covalently linked to RNA in the two VPgs were, surprisingly, not conserved; it is a tyrosine at position 5 of CfMV VPg and serine at position 1 of RYMV VPg. Phosphorylations were identified in CfMV and RYMV VPgs with two positionally similar locations T20/S14 and S71/S72, respectively. RYMV VPg contains an additional phosphorylation site at S41. INTRODUCTIONCocksfoot mottle virus (CfMV) and Rice yellow mottle virus (RYMV) are members of the genus Sobemovirus, a group of viruses with small icosahedral virions and a positive-sense ssRNA genome of approximately 4.0-4.5 kb. Like many other genera with an RNA genome, sobemoviruses have a viral genome-linked protein (VPg) attached to the 59 end of the genomic and subgenomic RNAs (Ghosh et al., 1981;Mang et al., 1982).The VPgs of sobemoviruses are translated as part of the polyprotein and cleaved by the viral protease (Nair & Savithri, 2010;van der Wilk et al., 1998). In contrast to potyviruses, the polyprotein processing and VPg maturation of sobemoviruses is poorly described. The specificity of the sobemoviral protease has been proposed as Q, E/T, S, N (Gorbalenya et al., 1988; Mäkinen et al., 2000;Nair & Savithri, 2010;van der Wilk et al., 1998), based on the fact that many different cleavage sites can be predicted for the N and C termini of sobemovirus VPgs. For several sobemoviruses -CfMV, RYMV, Southern bean mosaic virus (SBMV) and Sesbania mosaic virus (SeMV) -the N terminus of VPg has been mapped (Hébrard et al., 2008; Mäkinen et al., 2000;Nair & Savithri, 2010;van der Wilk et al., 1998), while the C terminus of VPg has so far been experimentally proven for only SeMV (Nair & Savithri, 2010). The determined SeMV VPg processing sites corroborate the predicted consensus cleavage sequence. However, sobemoviruses deploy 21 programmed ribosomal frameshifting (21 PRF) for the expression of polyprotein and VPg occupies a position in the polyprotein close to the 21 PRF signal. Therefore, it has been proposed that at least CfMV might express its VPg through the 21 PRF mechanism and as a result even encode VPgs with different C termini (Mäkinen et al., 2000).The VPgs are covalently linked to the 59 end of viral RNA (Ambros & Baltimore, 1978;Rothberg et al., 1978). The VPg is attached to the RNA over a phosphodiester bond formed between the hydroxyl group of the amino acid residue and 59 phosphate group of RNA (Ambros & Baltimore, 1978;Rothberg et al., 1978). The amino acid residue involved in the linkage has been reported to be a tyrosine or a serine (Ambros & Baltimore, 19...

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Binding analyses for the interaction between plant virus genome-linked protein (VPg) and plant translational initiation factors

Cited by 73 publications

References 70 publications

Functional Dissection of Naturally Occurring Amino Acid Substitutions in eIF4E That Confers Recessive Potyvirus Resistance in Plants

Functional Dissection of Naturally Occurring Amino Acid Substitutions in eIF4E That Confers Recessive Potyvirus Resistance in Plants

Visualization of the Interaction between the Precursors of VPg, the Viral Protein Linked to the Genome ofTurnip Mosaic Virus, and the Translation Eukaryotic Initiation Factor iso 4E In Planta

Protein-RNA linkage and post-translational modifications of two sobemovirus VPgs

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