HTLV-1 Tax Disrupts DNA Damage Repair-Response Complexes

描述（由申请人提供）：人类t细胞白血病病毒1型引起成人t细胞白血病和htlv相关脊髓病/热带痉挛性截瘫。据信，受感染的t细胞被病毒蛋白Tax转化导致疾病。税收介导细胞转化的过程尚不清楚，但被认为涉及税收介导的细胞基因组不稳定性。这些研究的目的是揭示税收破坏正常细胞DNA损伤识别-修复反应的机制，从而促进基因组的不稳定性。我们特别假设，Tax靶向并在功能上破坏调节细胞DNA损伤反应、修复识别和检查点激活关键方面的核复合体。我们提出（并将证实），Tax靶向的核复合体与一般文献中描述的DNA损伤形成的“损伤病灶”相同。我们将使用活细胞反卷积和共聚焦显微镜来确定Tax靶向这种复合物的机制。含税复合物的成分组成和物理特性将使用基于复杂质谱法的方法确定。Tax对这些复合物的结构和功能影响将通过定量蛋白质组学和分子功能分析的结合来揭示。本提案中所描述的研究的成功完成将提供一种机制，通过表达导致基因组不稳定。阐明这一因果模型将有助于我们对htlv -1介导的疾病的理解。

DESCRIPTION (provided by applicant): The Human T-Cell Leukemia Virus type 1 causes Adult T-Cell Leukemia and HTLV-Associated Myelopathy/Tropical Spastic Paraparesis. It is believed that transformation of the infected T-cell by the viral protein Tax results in disease. The process by which Tax mediates cellular transformation is not well understood but is thought to involve Tax-mediated cellular genomic instability. The objective of these studies is to uncover the mechanism by which Tax disrupts normal cellular DNA damage recognition-repair response, thus promoting genomic instability. We specifically hypothesize that Tax targets to and functionally disrupts a nuclear complex that regulates key aspects of cellular DNA damage response, repair recognition and checkpoint activation. We propose (and will confirm) that the nuclear complex targeted by Tax is the same "damage foci" described in the general literature as forming in response to DNA damage. We will use live-cell deconvolution and confocal microscopy to define the mechanism by which Tax targets this complex. The component make-up and physical characteristics of the Tax-containing complexes will be determined using sophisticated mass spectrometry based approaches. The structural and functional impact of Tax on these complexes will be revealed by a combination of quantitative proteomics and molecular functional assays. The successful completion of the studies described in this proposal will provide a mechanism by which expression of Tax results in genomic instability. Elucidation of this cause and effect model will contribute significantly to our understanding of HTLV-1-mediated disease.