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Structural and functional analyses of the FAM46 proteins.

FAM46 蛋白的结构和功能分析。

基本信息

批准号：
10334419
负责人：
Xuewu Zhang
金额：
$ 36.32万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-08 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10334419
关键词：
Aneuploidy BRCA2 gene Binding Biochemical Biological Assay Biophysics CDKN1A gene Cell Cycle Cell Cycle Stage Cell Nucleus Cell physiology Cells Centrosome Chromosome Segregation Cilia Clinic Complex Crystallization Cytoplasm DNA Damage DNA Repair Data Development Disease Ensure Family Family member Fluorescence Fluorescence Microscopy Genes Goals Homeostasis Hyperactivity Interphase Cell Knowledge Link Malignant Neoplasms Mediating Microtubules Mitotic spindle Molecular Multiple Myeloma Mutate Mutation Mutation Analysis Nuclear Organelles Organogenesis PLK1 gene Pathologic Patients Physiological Play Process Protein Analysis Proteins Regulation Resolution Roentgen Rays Role Signal Pathway Signal Transduction Structure Testing Therapeutic Tissues Work base biophysical techniques cilium biogenesis human disease insight kinetosome new therapeutic target novel nucleotidyltransferase protein protein interaction small molecular inhibitor

项目摘要

FAM46C (Family with sequence similarity 46, C) is one of the most frequently mutated genes in multiple myeloma (found in over 20% of the patients). Mutations of other FAM46 family members (A, B and D) are also associated with various human diseases. Despite the strong connections with diseases, the functions of FAM46 in either physiological or pathological settings are unknown. The goal of the project is to fill the knowledge gap on the functions of the FAM46 proteins and the underlying mechanisms, paving the way for developing therapeutic strategies for the associated diseases. The project is based on our preliminary work showing the direct physical interactions of FAM46 with polo-like kinase 4 (Plk4) and BRCA2 and CDKN1A(p21) interacting protein (BCCIP). Plk4 is the master regulator of centrosome duplication and ciliogenesis. BCCIP has been shown to be involved in regulating DNA repair and centrosome duplication. These together suggest that the FAM46 proteins regulate centrosome duplication, ciliogenesis and DNA repair. Centrosomes organize the bi-polar spindle formation and proper chromosome segregation in the cell cycle. In non-dividing cell, centrosomes mediate the formation of primary cilia, specialized signaling organelles critical for organogenesis and tissue homeostasis. A role in regulating these processes are consistent with the strong connection between FAM46 mutations and cancer. These hypotheses will be tested in three aims by combining X-ray crystallographic, biophysical, biochemical and cell-based functional approaches. Aim 1 will be focused on biophysical and structural analyses of the FAM46/Plk4 interaction. These studies will reveal the binding mode between FAM46 and Plk4, and identify key residues that can be tested by mutations in functional assays. In addition, potential mutual regulation of the enzymatic activities between FAM46 and Plk4 will be analyzed. Aim 2 will be focused on biophysical and structural analyses of the FAM46/BCCIP interaction. The crystal structure of the FAM46 and BCCIP complex will be determined to elucidate their interaction in atomic detail. Structural analyses will also be directed at the potential FAM46, Plk4 and BCCIPα tripartite complex. Mutational analyses will follow to test the binding mode and interface residues. The goal of Aim 3 is to analyze the roles of the interactions among FAM46, Plk4 and BCCIPα in regulating centrosome duplication, ciliogenesis and DNA repair. Fluorescence microscopy will be used to probe the localization of FAM46, Plk4 and BCCIPα, and their colocalization in and out of centrosomes in various stages of the cell cycle. The regulation of centrosome duplication, ciliogenesis and DNA repair by FAM46 through the interactions with Plk4 and BCCIP will also be analyzed by fluorescence microscopy.

FAM46C(具有序列相似性的家族46，C)是多发性骨髓瘤中最常见的突变基因之一。骨髓瘤(发现于20%以上的患者)。其他FAM46家族成员(A、B和D)的突变也是与各种人类疾病有关。尽管与疾病有很强的联系，但它的功能 FAM46在生理或病理环境中的作用尚不清楚。该项目的目标是填补对FAM46蛋白的功能和潜在机制的知识差距，为制定相关疾病的治疗策略。这个项目是以我们的前期工作为基础的 FAM46与Polo-like kinase4(Plk4)、BRCA2和CDKN1A(P21)的直接物理相互作用相互作用蛋白(BCCIP)Plk4是中心体复制和纤毛发生的主要调节因子。国商银行已被证明参与调节DNA修复和中心体复制。这些加在一起表明 FAM46蛋白调控中心体复制、纤毛发生和DNA修复。中心体组织细胞周期中两极纺锤体的形成和适当的染色体分离。在非分割单元中，中心体介导初级纤毛的形成，初级纤毛是器官发生中至关重要的特殊信号细胞器。和组织动态平衡。在调节这些过程中的作用与强联系是一致的 FAM46突变与癌症之间的关系。这些假设将通过结合X射线在三个目标上进行检验结晶学、生物物理学、生物化学和基于细胞的功能方法。目标1将专注于 FAM46/Plk4相互作用的生物物理和结构分析。这些研究将揭示绑定模式在FAM46和Plk4之间，并确定可以通过功能分析中的突变来检测的关键残基。在……里面此外，还将分析FAM46和Plk4之间酶活性的潜在相互调节。目标 2将侧重于FAM46/BCCIP相互作用的生物物理和结构分析。晶体结构将确定FAM46和BCCIP复合体的原子细节，以阐明它们的相互作用。结构性分析还将针对潜在的FAM46、Plk4和BCCIPα三方复合体。突变分析随后将测试结合模式和界面残留量。目标3的目标是分析 FAM46、Plk4和BCCIPα在调节中心体复制、纤毛发生和α中的相互作用修理。将利用荧光显微镜来探测FAM46、Plk4和BCCIPα的定位，以及它们的在细胞周期的不同阶段，中心体内外的共定位。中心体的调控 FAM46通过与Plk4和BCCIP的相互作用复制、纤毛发生和DNA修复也将是通过荧光显微镜进行分析。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Inhibition of FAM46/TENT5 activity by BCCIPα adopting a unique fold.

通过BCCIPα抑制FAM46/TENT5活性，采用独特的折叠。

DOI：
10.1126/sciadv.adf5583
发表时间：
2023-04-05
期刊：
SCIENCE ADVANCES
影响因子：
13.6
作者：
Liu, Shun;Chen, Hua;Yin, Yan;Lu, Defen;Gao, Guoming;Li, Jie;Bai, Xiao-Chen;Zhang, Xuewu
通讯作者：
Zhang, Xuewu

Cryo-EM structure of the Hippo signaling integrator human STRIPAK.