Proteomic profiling in Drosophila reveals potential Dube3a regulation of the actin cytoskeleton and neuronal homeostasis.
Proteomic profiling in Drosophila reveals potential Dube3a regulation of the actin cytoskeleton and neuronal homeostasis.
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DOI：
10.1371/journal.pone.0061952
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Reiter LT
中科院分区：
文献类型：
--
作者：
Jensen L;Farook MF;Reiter LT
Angelman综合征（AS）和15q重复自闭症相关的分子缺陷与E3泛素连接酶蛋白UBE3A的表达水平直接相关。在这里，我们使用黑腹果蝇在果蝇Dube3a或人类UBE3A蛋白水平降低（如as）或增加（如dup(15)）的条件下筛选这种泛素连接酶的靶标。利用全蝇头提取物的液相等电聚焦，我们确定了总共50种蛋白质，当Dube3a波动时，蛋白质和某些情况下的转录水平会发生变化。我们分析了细胞质，核和膜部分的头部提取物中Dube3a调节蛋白的含量。我们的研究结果表明，Dube3a参与了与ATP合成/代谢、肌动蛋白细胞骨架完整性、分解代谢和碳水化合物代谢以及神经系统发育和功能相关的细胞功能的调节。62%的蛋白质与同源的人类蛋白质相同，其中8种先前已被证明在苍蝇神经系统中泛素化。8种蛋白可能在转录水平上通过Dube3a的转录共激活功能受到Dube3a的调控。我们研究了一种自闭症相关蛋白ATPα，发现它可以以Dube3a依赖的方式泛素化。我们还发现Dube3a突变体的丝状肌动蛋白明显少于野生型幼虫，这与Dube3a调节肌动蛋白靶点的鉴定一致。UBE3A靶点的鉴定是揭示AS和重复15q自闭症分子病因学的第一步。
The molecular defects associated with Angelman syndrome (AS) and 15q duplication autism are directly correlated to expression levels of the E3 ubiquitin ligase protein UBE3A. Here we used Drosophila melanogaster to screen for the targets of this ubiquitin ligase under conditions of both decreased (as in AS) or increased (as in dup(15)) levels of the fly Dube3a or human UBE3A proteins. Using liquid phase isoelectric focusing of proteins from whole fly head extracts we identified a total of 50 proteins that show changes in protein, and in some cases transcriptional levels, when Dube3a fluctuates. We analyzed head extracts from cytoplasmic, nuclear and membrane fractions for Dube3a regulated proteins. Our results indicate that Dube3a is involved in the regulation of cellular functions related to ATP synthesis/metabolism, actin cytoskeletal integrity, both catabolism and carbohydrate metabolism as well as nervous system development and function. Sixty-two percent of the proteins were >50% identical to homologous human proteins and 8 have previously be shown to be ubiquitinated in the fly nervous system. Eight proteins may be regulated by Dube3a at the transcript level through the transcriptional co-activation function of Dube3a. We investigated one autism-associated protein, ATPα, and found that it can be ubiquitinated in a Dube3a dependent manner. We also found that Dube3a mutants have significantly less filamentous actin than wild type larvae consistent with the identification of actin targets regulated by Dube3a. The identification of UBE3A targets is the first step in unraveling the molecular etiology of AS and duplication 15q autism.
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影响因子：
8.8
作者：
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