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

摘要 染色质重塑复合体中的平行对数转换在调控的发生和丧失中起关键作用 对染色质旁对数功能的控制与多种癌症密切相关。人类SIN3A和SIN3B 是这类重要的染色质重塑的同源基因。SIN3A突变被认为是驱动因素 子宫体子宫内膜癌基因突变与SIN3A和SIN3B单倍体缺陷相关 人类神经发育障碍。人类SIN3A和SIN3B是几个大的多蛋白的成员 组蛋白脱乙酰酶HDAC1和HDAC2参与染色质重塑的复合体。 虽然SIN3A和SIN3B复合体的总体成员大多已知，但SIN3A和SIN3B复合体的结构和功能 这些网络中的特定复合体仍然知之甚少。此外，基因的精确靶向 SIN3/HDAC介导组蛋白脱乙酰化的抑制作用可能由非催化的SIN3亚基控制 但也仍然是个谜。这些关于SIN3A结构和功能的基本知识差距 含有SIN3B的复合体限制了我们设计癌症特定治疗干预措施的能力， 举个例子。实验室研究的首要目标是了解其分子机制。 特异的SIN3A和SIN3B复合体如何调节正常和 病态的人类状况。我们处于独特的地位，可以推动对这些研究领域的理解 基于我在染色质重塑和蛋白质组学方面的良好记录和长期专业知识， 合作者组成的团队，以及多学科的方法。在我们的实验室里，我们不断开发新的 蛋白质组学技术，并将其应用于染色质重塑复合体和网络。在这里，我们将使用 亲和纯化、酶动力学、多肽合成、荧光显微镜、 基因组学、定量蛋白质组学、交联质谱学和综合结构建模。vbl.使用 在接下来的五年里，我们将调查1)SIN3A和SIN3B复合体是如何 组装，2)特定亚基的调节功能，3)SIN3A复合体如何调节 特异性转录因子功能，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体中的蛋白质相互作用？这组问题将从一个 这些络合物的基本结构和功能决定了特定络合物在重要的 人体细胞模型。我们将为解决多样性和功能方面的知识差距奠定基础 特定的染色质重塑复合体。此外，我们将开始开发新的治疗策略，以 靶向特定的染色质重塑复合体。有了这些信息，我们将更接近我们的长期 设计基于多肽的方法来破坏特定蛋白质复合体的目标和愿景 特定的癌症治疗药物。

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.