RecQ DNA Helicase Impact on the Nuclear Pore Complex in Aging Cells

描述（由申请人提供）：作为基因组的守护者，RecQ解旋酶保存我们的遗传编码，从而保护我们的细胞免受衰老。同样，核孔复合物（NPCs）通过蛋白质和mRNA的转运维持适当的核功能。根据我们的初步工作，我们假设在RecQ解旋酶和NPC中的特定蛋白质之间存在联系，这些蛋白质共同起着维持基因组寿命的作用。我们的目标是通过靶向蛋白质组学收集定量数据来证明这种相互依赖性。这些方法的应用将识别重要的RecQ解旋酶和nup。我们建议1)进一步测试敲除RecQ解螺旋酶的DT40细胞和突变的RecQ解螺旋酶成纤维细胞Nup蛋白和mRNA水平，2)通过监测NLS-GFP的能量梯度和转运来分析核转运的变化，3)通过靶向蛋白质组学获得NPC化学计量的精确测量。

DESCRIPTION (provided by applicant): As guardians of the genome, RecQ helicases preserve our genetic coding and therefore protect our cells against aging. Likewise, Nuclear Pore Complexes (NPCs) maintain proper nuclear functionality through the transport of protein and mRNA. We hypothesize, from our preliminary work, that there exists a connection between RecQ helicases and specific proteins in the NPC that function together in maintenance of genomic longevity. Our goal is to gather quantitative data through targeted proteomics to prove this interdependence. Application of these methods will identify important RecQ helicases and Nups. We propose 1) to further test a panel of RecQ helicase knockout DT40 cells and mutant RecQ helicase fibroblasts for Nup protein and mRNA levels, 2) to analyze changes in nuclear transport by monitoring energy gradients and transport of NLS-GFP, 3) to obtain accurate measurements of NPC stoichiometry through targeted proteomics.