DIFFERENTIAL ANALYSIS OF WRN INTERACTING PARTNERS AFTER CISPLATINUM TREATMENT

这个子项目是许多利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，但不一定是研究者所在的机构。 已知缺乏Werner蛋白（WRN）的细胞对顺铂（cis-Pt）特别敏感，顺铂是一种通过交联DNA并引发细胞凋亡起作用的化疗药物。我们对暴露于这种药物的细胞的分析旨在了解细胞用于解决所产生的损伤的机制，以及WRN蛋白如何参与这些机制。 为了了解WRN参与，我们的目标是分析其相关的蛋白质复合物。通过使用遗传编码的TAP标签，我们正在从人胚肾（HEK-293）细胞的全细胞裂解物中富集WRN及其伴侣，并建议通过质谱分析这些复合物。 我们的实验室先前已经建立了用这些化学治疗化合物处理细胞的条件，并且已经进行了细胞处理的初步实验。我们正在等待第一个MS结果。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. It is known that cells deficient in the Werner protein (WRN) are specifically sensitive to cisplatinum (cis-Pt), a chemotherapy drug that works by crosslinking DNA and triggering apoptosis. Our profiling analysis of cells exposed to this drug is aimed at understanding the mechanisms cells employ to resolve the resulting damage, and how WRN protein is involved in these mechanisms. In order to understand WRN involvement, we aim to analyze its associated protein complex. By using a genetically encoded TAP tag, we are enriching for the WRN and its partners from whole cell lysates in human embryonic kidney (HEK-293) cells and are proposing to analyze these complexes by profiling mass spectrometry. Our lab has previously established conditions for treating cells with these chemotherapeutic compounds, and initial experiments with cell treatment have been performed. We are currently awaiting first MS results.