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

摘要 染色质重塑复合物中的Parkinson转换在调节性细胞凋亡的发生和丧失中起着关键作用。 对染色质旁核功能的控制与许多类型的癌症密切相关。人SIN 3A和SIN 3B 属于这类重要的染色质重塑旁系同源物。SIN 3A突变被认为是 在子宫体子宫内膜癌中的突变，以及SIN 3A和SIN 3B单倍不足与子宫内膜癌相关。 人类神经发育障碍人SIN 3A和SIN 3B是几个大的多蛋白的成员 参与染色质重塑的复合物，含有组蛋白脱乙酰酶HDAC 1和HDAC 2。 虽然SIN 3A和SIN 3B复合物的全部成员在很大程度上是已知的，但是SIN 3A和SIN 3B复合物的结构和功能是未知的。 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 SIN 3/HDAC介导的组蛋白去乙酰化的阻遏可能是由非催化性SIN 3亚基控制的 但也是个谜这些关于SIN 3A结构和功能的基本知识空白 和含SIN 3B的复合物限制了我们设计癌症特异性治疗干预的能力， example.实验室研究的首要目标是了解 特异性SIN 3A和SIN 3B复合物如何调节正常和 病态的人类状况我们处于一个独特的位置，以促进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， 集合合作者团队和多学科方法。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物和网络。在这里，我们将使用 亲和纯化，酶动力学，肽合成，荧光显微镜， 基因组学、定量蛋白质组学、交联质谱和综合结构建模。使用 这些方法在未来五年，我们将研究1）如何SIN 3A和SIN 3B复合物 2）特定亚基的调节功能，3）SIN 3A复合物如何调节 特异性转录因子功能，4）子宫内膜细胞中的SIN 3A和SIN 3B网络是什么，以及5） 我们如何破坏这些复合物中的蛋白质相互作用？这组问题将从一个 这些复合物的基本结构和功能，以确定特定复合物的作用， 人类细胞模型我们将为解决知识在多样性和功能上的差距奠定基础 特定的染色质重塑复合物。此外，我们将开始开发新的治疗策略， 靶向特异性染色质重塑复合物。有了这些信息，我们将更接近我们的长期目标。 设计基于肽的方法来破坏特定蛋白质复合物的目标和愿景是新颖的， 特异性癌症治疗剂。

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.