Article

Coupling Genetics and Proteomics To Identify Aphid Proteins Associated with Vector-Specific Transmission of Polerovirus (Luteoviridae)

Department of Plant Pathology, Cornell University, Ithaca, NY 14853, USA.
Journal of Virology (Impact Factor: 4.65). 02/2008; 82(1):291-9. DOI: 10.1128/JVI.01736-07
Source: PubMed

ABSTRACT Cereal yellow dwarf virus-RPV (CYDV-RPV) is transmitted specifically by the aphids Rhopalosiphum padi and Schizaphis graminum in a circulative nonpropagative manner. The high level of vector specificity results from the vector aphids having the functional components of the receptor-mediated endocytotic pathways to allow virus to transverse the gut and salivary tissues. Studies of F(2) progeny from crosses of vector and nonvector genotypes of S. graminum showed that virus transmission efficiency is a heritable trait regulated by multiple genes acting in an additive fashion and that gut- and salivary gland-associated factors are not genetically linked. Utilizing two-dimensional difference gel electrophoresis to compare the proteomes of vector and nonvector parental and F(2) genotypes, four aphid proteins (S4, S8, S29, and S405) were specifically associated with the ability of S. graminum to transmit CYDV-RPV. The four proteins were coimmunoprecipitated with purified RPV, indicating that the aphid proteins are capable of binding to virus. Analysis by mass spectrometry identified S4 as a luciferase and S29 as a cyclophilin, both of which have been implicated in macromolecular transport. Proteins S8 and S405 were not identified from available databases. Study of this unique genetic system coupled with proteomic analysis indicated that these four virus-binding aphid proteins were specifically inherited and conserved in different generations of vector genotypes and suggests that they play a major role in regulating polerovirus transmission.

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    • "Acetyl-CoA is a key molecule linking cellular metabolism and protein function via acetylation in plants (Xing & Poirier, 2012). Quite intriguingly, a major protein differentially expressed in both lab-reared and naturally occurring vector and non-vector populations of Schizaphis graminum, a vector species for CYDV-RPV, is an acetyl- CoA ligase (Cilia et al., 2011a, b; Yang et al., 2008). Those data suggest that S. graminum CoA ligase may play a central role in determining the availability of acetyl groups to viral or aphid protein substrates and that regulation of the acetylation helps to determine variation in vectoring capacity in this species. "
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    ABSTRACT: Virions of the RPV strain of Cereal yellow dwarf virus (CYDV-RPV) were purified from infected oat tissue and analyzed by mass spectrometry. Two conserved residues, K147 and K181, residing in the virus coat protein, were confidently identified to contain epsilon-N-acetyl groups. While no functional data are available for K147, K181 lies within an interfacial region critical for virion assembly and stability. The signature immonium ion at m/z 126.0919 demonstrated the presence of N-acetyllysine, and the sequence fragment ions enabled an unambiguous assignment of the epsilon-N-acetyl modification on K181. We hypothesize that selection favors acetylation of K181 in a fraction of coat protein monomers to stabilize the capsid by promoting intermonomer salt bridge formation.
    Journal of General Virology 06/2014; 95(Pt 10). DOI:10.1099/vir.0.066514-0 · 3.53 Impact Factor
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    • "The identification of a single protein assumed to be S29 as interacting with CYDV-RPV in the Yang et al. (2008) experiment conflicts with the his-tagging experiment (Figure 2) that showed both S28 and S29 interact with CYDV-RPV. Our co-IP experiment with aphid and CYDV- RPV indicates the potential involvement of a third cyclophilin protein, cyclophilin A. One hypothesis is that the protein spot from the co-IP-DIGE reported by Yang et al. 2008 contained cyclophilin A and not either of the two cyclophilin B isoforms or perhaps a mixture of both. The difference in results could be explained by the differences in techniques used and the detection limits of each analytical platform. "
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    ABSTRACT: Yellow dwarf viruses cause the most economically important virus diseases of cereal crops worldwide and are transmitted by aphid vectors. The identification of aphid genes and proteins mediating virus transmission is critical to develop agriculturally sustainable virus management practices and to understand viral strategies for circulative movement in all insect vectors. Two cyclophilin B proteins, S28 and S29, were identified previously in populations of Schizaphisgraminum that differed in their ability to transmit the RPV strain of Cereal yellow dwarf virus (CYDV-RPV). The presence of S29 was correlated with F2 genotypes that were efficient virus transmitters. The present study revealed the two proteins were isoforms, and a single amino acid change distinguished S28 and S29. The distribution of the two alleles was determined in 12 F2 genotypes segregating for CYDV-RPV transmission capacity and in 11 genetically independent, field-collected S. graminum biotypes. Transmission efficiency for CYDV-RPV was determined in all genotypes and biotypes. The S29 isoform was present in all genotypes or biotypes that efficiently transmit CYDV-RPV and more specifically in genotypes that efficiently transport virus across the hindgut. We confirmed a direct interaction between CYDV-RPV and both S28 and S29 using purified virus and bacterially expressed, his-tagged S28 and S29 proteins. Importantly, S29 failed to interact with a closely related virus that is transported across the aphid midgut. We tested for in vivo interactions using an aphid-virus co-immunoprecipitation strategy coupled with a bottom-up LC-MS/MS analysis using a Q Exactive mass spectrometer. This analysis enabled us to identify a third cyclophilin protein, cyclophilin A, interacting directly or in complex with purified CYDV-RPV. Taken together, these data provide evidence that both cyclophilin A and B interact with CYDV-RPV, and these interactions may be important but not sufficient to mediate virus transport from the hindgut lumen into the hemocoel.
    PLoS ONE 08/2013; 8(8):e71620. DOI:10.1371/journal.pone.0071620 · 3.53 Impact Factor
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    • "the salivary duct, from which they can infect potential host plants (Gildow, 1985, 1993; Gildow and Gray, 1993; Yang et al., 2008). Virus-aphid specificity likely results from the recognition between virions of a specific isolate and the viral receptors in the accessory salivary glands of a particular aphid species (Gildow and Rochow, 1980; Gildow and Gray, 1993; Peiffer et al., 1997; Bencharki et al., 2000). "
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    Journal of virological methods 07/2013; 194(1-2). DOI:10.1016/j.jviromet.2013.07.038 · 1.88 Impact Factor
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