Genome-wide annotation and functional identification of aphid GLUT-like sugar transporters.
Genome-wide annotation and functional identification of aphid GLUT-like sugar transporters.
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DOI：
10.1186/1471-2164-15-647
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发表时间：
2014-08-04
期刊：
影响因子：
4.4
通讯作者：
Gatehouse JA
中科院分区：
文献类型：
--
作者：
Price DR;Gatehouse JA
以韧皮部为食的昆虫，如蚜虫，几乎连续地以植物韧皮部汁液为食，这是一种含有高浓度蔗糖（一种由葡萄糖和果糖组成的二糖）的液体食物。为了获得可用的碳，蚜虫在肠腔中水解蔗糖并运输其组成单糖，葡萄糖和果糖。虽然糖运输在蚜虫营养中起着至关重要的作用，但蚜虫中糖运输的分子基础，以及更普遍的所有昆虫中的糖运输的分子基础，仍然缺乏特征。在这里，使用最新发布的豌豆蚜虫，Acyrthosiphon豌豆，基因组，我们提供了一个更新的基因注释和表达谱的推定糖转运蛋白。最后，在酵母中功能性表达肠道表达的糖转运蛋白，并筛选葡萄糖和果糖转运活性。在这项研究中，我们使用从头开始的方法，确定了19糖转运蛋白（SP）家族转运蛋白在A。豌豆基因组基因表达分析，基于214，834 A. pisum表达序列标签，支持17个糖转运蛋白家族转运蛋白在成年雌性蚜虫中活跃表达。进一步分析，采用定量PCR鉴定了4种转运蛋白，A.豌豆糖转运蛋白1、3、4和9（ApST 1、ApST 3、ApST 4和ApST 9）在肠组织中高度表达和/或富集。当在酿酒酵母己糖转运蛋白缺失突变体（菌株EBY.VW4000）中表达时，仅ApST 3（先前表征）和ApST 4（本文报道）转运葡萄糖和果糖，导致酵母突变体的功能性拯救。在这里，我们的特点ApST 4，491个氨基酸的蛋白质，与12个预测的跨膜区域，作为一个促进葡萄糖/果糖转运蛋白。最后，系统发育重建表明，ApST 4，和相关的，尚未表征的昆虫转运蛋白在遗传学上与人类GLUT（SLC 2A）I类促进葡萄糖/果糖转运蛋白密切相关。肠道增强ApST 4的表达，其产物的转运特异性与ApST 4作为肠道葡萄糖/果糖转运蛋白的功能一致。在这里，我们假设ApST 3（以前报道过）和ApST 4（这里报道过）都在肠道界面起作用，从肠腔输入葡萄糖和果糖。本文的在线版本（doi：10.1186/1471-2164-15-647）包含补充材料，可供授权用户使用。
Phloem feeding insects, such as aphids, feed almost continuously on plant phloem sap, a liquid diet that contains high concentrations of sucrose (a disaccharide comprising of glucose and fructose). To access the available carbon, aphids hydrolyze sucrose in the gut lumen and transport its constituent monosaccharides, glucose and fructose. Although sugar transport plays a critical role in aphid nutrition, the molecular basis of sugar transport in aphids, and more generally across all insects, remains poorly characterized. Here, using the latest release of the pea aphid, Acyrthosiphon pisum, genome we provide an updated gene annotation and expression profile of putative sugar transporters. Finally, gut expressed sugar transporters are functionally expressed in yeast and screened for glucose and fructose transport activity. In this study, using a de novo approach, we identified 19 sugar porter (SP) family transporters in the A. pisum genome. Gene expression analysis, based on 214, 834 A. pisum expressed sequence tags, supports 17 sugar porter family transporters being actively expressed in adult female aphids. Further analysis, using quantitative PCR identifies 4 transporters, A. pisum sugar transporter 1, 3, 4 and 9 (ApST1, ApST3, ApST4 and ApST9) as highly expressed and/or enriched in gut tissue. When expressed in a Saccharomyces cerevisiae hexose transporter deletion mutant (strain EBY.VW4000), only ApST3 (previously characterized) and ApST4 (reported here) transport glucose and fructose resulting in functional rescue of the yeast mutant. Here we characterize ApST4, a 491 amino acid protein, with 12 predicted transmembrane regions, as a facilitative glucose/fructose transporter. Finally, phylogenetic reconstruction reveals that ApST4, and related, as yet uncharacterized insect transporters are phylogenetically closely related to human GLUT (SLC2A) class I facilitative glucose/fructose transporters. The gut enhanced expression of ApST4, and the transport specificity of its product is consistent with ApST4 functioning as a gut glucose/fructose transporter. Here, we hypothesize that both ApST3 (reported previously) and ApST4 (reported here) function at the gut interface to import glucose and fructose from the gut lumen. The online version of this article (doi:10.1186/1471-2164-15-647) contains supplementary material, which is available to authorized users.
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影响因子：
14.8
作者：
Gietz, R. Daniel;Schiestl, Robert H.
通讯作者：
Schiestl, Robert H.
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14.9
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