Impact of pathogenic missense mutations on the ARID domain of ARID1a

致病性错义突变对 ARID1a ARID 结构域的影响

• 批准号: 10654137
• 负责人: Hugo Sanabria
• 金额: $ 44.31万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-12 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10654137
• 关键词: ARID Domain; ARID1A gene; Affinity; Amino Acids; Binding; Bioinformatics; Biomedical Research; Biophysics; Breast; Cancer Patient; Cell Proliferation; Cholangiocarcinoma; Chromatin; Chromatin Remodeling Factor; Classification; Clinical Trials; Complex; Computer Models; DNA; DNA Binding; DNA Binding Domain; DNA Repair; DNA biosynthesis; DNA-Protein Interaction; Databases; Environment; Fluorescence; Fluorescence Resonance Energy Transfer; Fostering; Foundations; Frameshift Mutation; Future; Gene Expression; Genes; Genetic Transcription; Goals; Health; Health Benefit; Helix-Turn-Helix Motifs; Libraries; Malignant Neoplasms; Malignant neoplasm of ovary; Measures; Missense Mutation; Modeling; Molecular Mechanisms of Action; Mutate; Mutation; Names; Non-Small-Cell Lung Carcinoma; Nuclear; Outcome; Ovarian; Pancreas; Pathogenicity; Pathway interactions; Patients; Physical condensation; Positioning Attribute; Prevention; Program Development; Property; Protein Family; Proteins; Research; Research Training; Science; Solid Neoplasm; Spectrum Analysis; Structural Models; Structure; Structure-Activity Relationship; Sucrose; System; Talents; Testing; The Cancer Genome Atlas; Therapeutic; Tumor Suppressor Genes; Tumor Suppressor Proteins; Universities; Uterus; Work; alpha helix; anticancer research; biobank; biophysical analysis; career; career development; design; experience; experimental study; fighting; graduate student; improved; insight; interest; loss of function; loss of function mutation; member; mutant; novel therapeutics; personalized medicine; physical science; programs; protein folding; resistance mutation; screening; single molecule; temporal measurement; translational oncology; undergraduate student

SUMMARY The AT-rich interacting domain-containing protein 1a (ARID1a), also named BAF250a, p270, or hOSA1, is a vital component of the Switch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeling complex. ARID1a, as part of the SWI/SNF complex, is responsible for crucial nuclear activities, including regulating transcription, DNA synthesis, DNA damage repair, and cell proliferation. Consequently, ARID1a is classified as a tumor suppressor gene and it is frequently mutated in solid tumor malignancies cancers, amounting to ~6% of every cancer and ~45% of all ovarian cancers. Loss of function mutations in ARID1a due to frameshift, stop-gain, or missense mutations leads to dysregulation of many gene pathways, including the prevention of tumor suppressor activities. Although the loss of function is evident in stop-gain and frameshift mutations, the impact of pathogenic missense mutations is subtler and more difficult to understand or predict. We hypothesize that pathogenic missense mutations impact the protein stability, DNA binding affinity, and structural dynamics, perturbing its function. Specifically, we seek to understand the effects of pathogenic missense mutations in the ARID domain of ARID1a, which is responsible for its direct interaction with DNA. Therefore, the broad goal of this research program is to understand the effects of pathogenic missense mutations in the ARID domain of ARID1a. The long-term implications of this program are the development of a unique framework that will pave the way for directly probing identified pathogenic missense mutations for future guided patient-specific screening therapeutic approaches. Our proposed study integrates biophysical studies, computational approaches, and single-molecule spectroscopy to characterize the stability, DNA binding, and structural dynamics of the ARID domain. Our studies will give insights into the correlation between missense pathogenic mutations and the structure-dynamics- function relationship. The PI and our research team are uniquely positioned to pursue the following specific aims: (1) to determine the impact of pathogenic missense mutations on the stability of the ARID domain; (2) to determine the impact of pathogenic missense mutations in the binding affinity of ARID domain to DNA; and (3) to determine the changes in the structure and dynamics of ARID domain as perturbed by pathogenic missense mutations. This interdisciplinary project will engage undergraduates in research to foster their interest and career development in both physical sciences and cancer research. The expected outcome will be a structural model of ARID1a’s ARID domain interaction with DNA, a library of pathogenic mutations ranked by the impact on stability and affinity, a system to improve pathogenic predictors, and the foundation for developing novel therapeutics and personalized medicine in the fight against cancer.

总结 富含AT的相互作用结构域蛋白1a（ARID 1a），也称为BAF 250 a，p270或hOSA 1，是一种重要的 SWI/SNF染色质重塑复合物的组成部分。ARID 1a，作为 SWI/SNF复合物的一部分，负责重要的核活动，包括调节转录、DNA 合成、DNA损伤修复和细胞增殖。因此，ARID 1a被归类为肿瘤抑制因子 该基因在实体瘤恶性肿瘤中经常发生突变，占每种癌症的约6%， 约45%的卵巢癌。由于移码、停止增益或错义导致的ARID 1a功能缺失突变 突变导致许多基因通路的失调，包括肿瘤抑制活性的阻止。 虽然功能丧失在停止增益和移码突变中是明显的，但致病性错义的影响 突变是微妙的，更难以理解或预测。我们假设致病性错义 突变影响蛋白质稳定性、DNA结合亲和力和结构动力学，扰乱其功能。 具体来说，我们试图了解ARID 1a的ARID结构域中致病性错义突变的影响， 它负责与DNA的直接相互作用。因此，这项研究计划的主要目标是 了解ARID 1a的ARID结构域中致病性错义突变的影响。长期 该计划的意义在于开发一个独特的框架，为直接探测 确定了致病性错义突变，用于未来指导患者特异性筛查治疗方法。 我们提出的研究整合了生物物理学研究，计算方法和单分子 光谱来表征ARID结构域的稳定性、DNA结合和结构动力学。我们的研究 将深入了解错义致病突变与结构动力学之间的相关性， 函数关系PI和我们的研究团队处于独特的位置，以追求以下特定目标： (1)确定致病性错义突变对ARID结构域稳定性的影响;（2） 确定致病性错义突变对ARID结构域与DNA的结合亲和力的影响;和（3） 确定致病性错义干扰下ARID结构域的结构和动力学变化 突变。这个跨学科的项目将使本科生参与研究，以培养他们的兴趣和职业生涯 物理科学和癌症研究的发展。预期的结果将是一个结构模型 ARID 1a的ARID结构域与DNA的相互作用，一个致病性突变库， 稳定性和亲和力，一个系统，以改善致病预测，并为开发新的基础 治疗学和个体化医学在对抗癌症中的作用。