HTLV-1 Tax Disrupts DNA Damage Repair-Response Complexes

描述（由申请方提供）：人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/热带痉挛性轻瘫。 据信，病毒蛋白Tax对受感染的T细胞的转化导致疾病。 Tax介导细胞转化的过程尚未完全了解，但被认为涉及Tax介导的细胞基因组不稳定性。 这些研究的目的是揭示Tax破坏正常细胞DNA损伤修复反应，从而促进基因组不稳定性的机制。 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物。我们建议（并将确认），核复合物的目标税收是相同的“损伤灶”中描述的一般文献中形成的DNA损伤。 我们将使用活细胞去卷积和共聚焦显微镜来定义Tax针对这种复合物的机制。 含税复合物的组分组成和物理特性将使用基于复杂质谱法的方法来确定。 税收对这些复合物的结构和功能的影响将通过定量蛋白质组学和分子功能测定的结合来揭示。 本提案中描述的研究的成功完成将提供Tax表达导致基因组不稳定性的机制。 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.