Genetics Coupled to Quantitative Intact Proteomics Links Heritable Aphid and Endosymbiont Protein Expression to Circulative Polerovirus Transmission
Genetics Coupled to Quantitative Intact Proteomics Links Heritable Aphid and Endosymbiont Protein Expression to Circulative Polerovirus Transmission
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DOI：
10.1128/jvi.01504-10
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发表时间：
2011-03-01
影响因子：
5.4
通讯作者：
Gray, S.
中科院分区：
文献类型：
--
作者：
Cilia, M.;Tamborindeguy, C.;Gray, S.
黄矮病毒科的黄矮病毒是谷类作物黄矮病的病原体，它们各自通过不同种类的蚜虫以循环方式最有效地传播，这需要病毒与多种蚜虫蛋白相互作用。使用二维差异凝胶电泳 (DIGE) 与基质辅助激光解吸电离串联质谱或在线纳米级液相色谱联用，鉴定了 F2 禾谷裂霉基因型中差异表达的蚜虫蛋白，这些蛋白具有传播谷物黄矮病毒 RPV (CYDV-RPV) 的能力 与电喷雾串联质谱联用。总共 50 个蛋白质点，包含蚜虫蛋白质以及来自蚜虫专性和母系遗传的细菌内共生体 Buchnera 的蛋白质，被鉴定为在具有传播能力和折射能力的蚜虫之间存在差异表达。令人惊讶的是，在具有病毒传播能力的 F2 基因型中，Buchnera 蛋白的等电点与预期的母本 Buchnera 蛋白质组中的等电点不匹配，而是与具有传播能力的父本的 Buchnera 蛋白质组一致。在已鉴定的蚜虫蛋白中，许多与能量代谢、膜运输、脂质信号传导和细胞骨架有关。至少有八种蚜虫蛋白被表达为可遗传的等电点亚型对，每个亲本谱系都有一个亚型对。在F2基因型中，来自亲本谱系的蚜虫蛋白亚型的表达与病毒传播表型高度精确地一致。因此，这些同种型是禾谷菌中 CYDV-RPV 传播的候选生物标志物。我们结合遗传和 DIGE 方法还可以预测几种蛋白质在具有不同传播障碍的折射蚜虫中的表达位置。预计有 12 种蛋白质在蚜虫的后肠中起作用，而有 6 种蛋白质预计与附属唾液腺或血淋巴有关。了解调节病毒传播的蛋白质及其预测位置将有助于了解调节蚜虫循环病毒传播的生化机制，以及确定阻止传播的新靶标。
Yellow dwarf viruses in the family Luteoviridae, which are the causal agents of yellow dwarf disease in cereal crops, are each transmitted most efficiently by different species of aphids in a circulative manner that requires the virus to interact with a multitude of aphid proteins. Aphid proteins differentially expressed in F2 Schizaphis graminum genotypes segregating for the ability to transmit Cereal yellow dwarf virus-RPV (CYDV-RPV) were identified using two-dimensional difference gel electrophoresis (DIGE) coupled to either matrix-assisted laser desorption ionization-tandem mass spectrometry or online nanoscale liquid chromatography coupled to electrospray tandem mass spectrometry. A total of 50 protein spots, containing aphid proteins and proteins from the aphid's obligate and maternally inherited bacterial endosymbiont, Buchnera, were identified as differentially expressed between transmission-competent and refractive aphids. Surprisingly, in virus transmission-competent F2 genotypes, the isoelectric points of the Buchnera proteins did not match those in the maternal Buchnera proteome as expected, but instead they aligned with the Buchnera proteome of the transmission-competent paternal parent. Among the aphid proteins identified, many were involved in energy metabolism, membrane trafficking, lipid signaling, and the cytoskeleton. At least eight aphid proteins were expressed as heritable, isoelectric point isoform pairs, one derived from each parental lineage. In the F2 genotypes, the expression of aphid protein isoforms derived from the competent parental lineage aligned with the virus transmission phenotype with high precision. Thus, these isoforms are candidate biomarkers for CYDV-RPV transmission in S. graminum. Our combined genetic and DIGE approach also made it possible to predict where several of the proteins may be expressed in refractive aphids with different barriers to transmission. Twelve proteins were predicted to act in the hindgut of the aphid, while six proteins were predicted to be associated with the accessory salivary glands or hemolymph. Knowledge of the proteins that regulate virus transmission and their predicted locations will aid in understanding the biochemical mechanisms regulating circulative virus transmission in aphids, as well as in identifying new targets to block transmission.