Proteomic analysis of Mutator transposition

描述(申请人提供)：转座子是可以移动到基因组中新位置的遗传元件。转座子在几乎所有生物体中都普遍存在，最近在人类和玉米中的基因组测序项目表明，转座子代表了这两个基因组的很大一部分。在细胞水平上，转座子的活性需要DNA和染色体修复过程，以防止细胞损伤，这可能导致细胞不受控制的增殖。除了这些关于转座子活性后果的下游研究外，人们对体内调节其上游(切除和插入)行为的蛋白质知之甚少。这项研究将使用基于蛋白质组学的方法来识别调节转座行为的宿主蛋白质，并潜在地识别调控细胞增殖的候选蛋白质，以及建立一种可行的替代病毒载体的基因疗法。该项目的具体目标是：(1)比较Mu激活前后组织的蛋白质组；(2)提炼转座机制的空间表达模式；(3)挖掘基因组和cDNA数据库，以寻找移码翻译候选基因。与转座相关的蛋白质在四种选定的组织类型中的发育作用(S)将进一步利用分子、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.