Proteomic analysis of Mutator transposition

描述（由申请人提供）：转座子是可以移动到基因组新位置的遗传元件。转座子基本上在所有生物体中都存在，最近的人类和玉米基因组测序项目表明，转座子代表了两种基因组的重要部分。在细胞水平上，转座子的活性需要DNA和染色体修复过程来防止可能导致不受控制的细胞增殖的细胞损伤。除了这些关于转座子活性影响的下游研究外，对于调节转座子在体内上游（切除和插入）行为的蛋白质知之甚少。本研究将使用基于蛋白质组学的方法来鉴定调节转位行为的宿主蛋白，并可能鉴定调节细胞增殖的候选蛋白，以及建立一种可行的替代病毒载体的基因治疗方法。本项目的具体目的是：(1)比较Mu激活前后组织的蛋白质组学；(2)细化换位机制的空间表达模式；(3)挖掘基因组和cDNA数据库，鉴定移框翻译候选基因。在四种选定的组织类型中，与转位相关的蛋白质的发育作用将进一步使用分子，生化和反向遗传学方法进行分析。

DESCRIPTION (provided by applicant): Transposons are genetic elements that can move to new locations in the genome. They are ubiquitous in essentially all organisms, and recent genome sequencing projects in humans and maize have shown that transposons represent significant portions of both genomes. At the cellular level, transposon activity requires DNA and chromosomal repair processes to prevent cellular damage that could result in uncontrolled cell proliferation. Aside from these downstream studies on the consequences of transposon activity, relatively little is known about the proteins that modulate their upstream (excision and insertion) behavior in vivo. This study will use a proteomics-based approach to identify host proteins that regulate transposition behavior and potentially identify candidate proteins that modulate cell proliferation as well as establish a viable gene therapy alternative to viral vectors. The specific aims of this project are: (1) To compare the proteomes of tissues before and after activation of Mu; (2) to refine the spatial expression pattern of the transposition mechanisms; and (3) to mine genomic and cDNA databases to identify frame-shift translation candidate genes. The developmental role(s) of proteins associated with transposition in the four selected tissue types will be further analyzed using molecular, biochemical and reverse genetics approaches.