Decoding the Assembly and Function of Paralogue Specific SIN3A and SIN3B Human Chromatin Remodeling Complexes and Networks

解码旁系同源物特异性 SIN3A 和 SIN3B 人类染色质重塑复合物和网络的组装和功能

• 批准号: 10678855
• 负责人: MICHAEL P WASHBURN
• 金额: $ 38.75万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10678855
• 关键词: Address; Affinity Chromatography; Area; Body of uterus; Cell model; Cells; Chromatin; Chromatin Remodeling Factor; Complex; Development; Endometrial; Endometrial Carcinoma; Enzyme Kinetics; Enzymes; Fluorescence Microscopy; Foundations; Gene Expression; Genomics; Goals; HDAC1 gene; HDAC2 gene; Hand; Histone Deacetylase; Histone Deacetylation; Human; Knowledge; Laboratories; Link; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Methodology; Molecular; Multiprotein Complexes; Mutate; Mutation; Neurodevelopmental Disorder; Peptide Synthesis; Peptides; Play; Positioning Attribute; Proteomics; Public Health; Research; Role; Structural Models; Structure; Technology; Therapeutic Agents; Therapeutic Intervention; Vision; cancer type; chromatin remodeling; crosslink; design; driver mutation; gene repression; human disease; interdisciplinary approach; member; novel; novel therapeutic intervention; paralogous gene; protein complex; protein protein interaction; transcription factor

ABSTRACT Paralog switching in chromatin remodeling complexes plays a key role in development and loss of regulatory control over chromatin paralog functions is closely linked to many types of cancer. Human SIN3A and SIN3B are amongst this class of important chromatin remodeling paralogs. SIN3A mutations are considered driver mutations in uterine corpus endometrial carcinoma, and both SIN3A and SIN3B haploinsufficiency linked to human neurodevelopmental disorders. Human SIN3A and SIN3B are members of several large multiprotein complexes involved in chromatin remodeling containing the histone deacetylase enzymes HDAC1 and HDAC2. While the overall members of the SIN3A and SIN3B complexes is largely known, the structure and function of specific complexes in these networks remain poorly understood. Furthermore, the precise targeting of gene repression by SIN3/HDAC-mediated histone deacetylation is likely controlled by the non-catalytic SIN3 subunits but also remains enigmatic. These fundamental gaps in knowledge regarding the structure and function of SIN3A and SIN3B containing complexes limit our ability to devise specific therapeutic interventions in cancer, for example. The overarching goal of the research in the laboratory is to understand the molecular mechanisms of how specific SIN3A and SIN3B complexes regulate gene expression and chromatin remodeling in normal and diseased human conditions. We are in a unique position to advance the understanding of these area of research based on my strong track record and long-standing expertise in chromatin remodeling and proteomics, assembled team of collaborators, and multidisciplinary approach. In our lab, we continually develop new proteomic technologies and apply them to chromatin remodeling complexes and networks. Here, we will use a comprehensive approach of affinity purification, enzyme kinetics, peptide synthesis, fluorescence microscopy, genomics, quantitative proteomics, cross linking mass spectrometry, and integrative structural modeling. Using these methodologies over the next five years, we will investigate 1) How are SIN3A and SIN3B complexes assembled, 2) What are the regulatory functions of specific subunits, 3) How do SIN3A complexes regulate specific transcription factor function, 4) What are the SIN3A and SIN3B networks in endometrial cells, and 5) How can we disrupt protein protein interactions in these complexes? This set of questions will go from a fundamental structure and function of these complexes to determining the role of specific complexes in important human cellular models. We will lay the foundation to address the gap in knowledge in the diversity and function of specific chromatin remodeling complexes. In addition, we will begin to develop new therapeutic strategies to target specific chromatin remodeling complexes. With this information in hand, we will be closer to our long-term goal and vision of designing peptide-based approaches to disrupt specific protein complexes as novel and specific cancer therapeutic agents.

抽象的 谱系染色质重塑络合物中的旁系同源物转换在调节的发展和丢失中起关键作用 控制染色质旁同产功能与多种类型的癌症密切相关。人类sin3a和sin3b 是这类重要的染色质重塑旁系同源物之一。 SIN3A突变被认为是驾驶员 子宫内膜癌的突变，以及与 人类神经发育障碍。人类SIN3A和SIN3B是几种大型多蛋白的成员 含有组蛋白脱乙酰基酶HDAC1和HDAC2的染色质重塑的复合物。 虽然SIN3A和SIN3B复合物的总成员在很大程度上是已知的，但 这些网络中的特定复合物仍然知之甚少。此外，基因的精确靶向 SIN3/HDAC介导的组蛋白脱乙酰化的抑制可能受非催化SIN3亚基的控制 但也仍然是神秘的。关于SIN3A的结构和功能知识的这些基本差距 含有复合物的SIN3b限制了我们设计特定的癌症治疗干预措施的能力，因为 例子。该研究中研究的总体目标是了解 特定的SIN3A和SIN3B复合物如何调节基因表达和染色质重塑在正常和 患病的人类状况。我们处于一个独特的位置，可以促进对这些研究领域的理解 根据我的良好往绩和染色质重塑和蛋白质组学方面的长期专业知识， 组合的合作者团队和多学科方法。在我们的实验室中，我们不断发展新的 蛋白质组学技术并将其应用于染色质重塑复合物和网络。在这里，我们将使用 亲和力纯化，酶动力学，肽合成，荧光显微镜，综合方法 基因组学，定量蛋白质组学，交叉连接质谱和整合结构建模。使用 这些方法在未来五年内，我们将研究1）SIN3A和SIN3B复合物如何 组装，2）特定亚基的调节功能是什么，3）SIN3A复合物如何调节 特定的转录因子函数，4）子宫内膜细胞中的SIN3A和SIN3B网络是什么，5） 我们如何破坏这些复合物中的蛋白质蛋白质相互作用？这组问题将来自 这些复合物的基本结构和功能，以确定特定复合物在重要的复合物中的作用 人类细胞模型。我们将奠定基础，以解决多样性和功能知识的差距 特定的染色质重塑复合物。此外，我们将开始制定新的治疗策略 靶标特异性染色质重塑络合物。借助此信息，我们将更接近我们的长期 设计基于肽的方法以破坏特定蛋白质复合物为新颖的特定蛋白质复合物的目标和愿景 特定的癌症治疗剂。