The structural and functional consequences of FAT10 phosphorylation (C09*)

FAT10 磷酸化的结构和功能后果 (C09*)

• 批准号: 436149870
• 金额: --
• 依托单位: Lehrstuhl Biologie/Immunologie
• 依托单位国家: 德国
• 项目类别: Collaborative Research Centres
• 财政年份: 2020
• 资助国家: 德国
• 起止时间: 2019-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/436149870?language=en
• 关键词: structural; functional; consequences; FAT10; phosphorylation

The phosphorylation of ubiquitin has major consequences for its function in mitophagy. Another modifier of the ubiquitin family, designated HLA-F adjacent transcript 10 (FAT10), that also targets conjugated proteins for degradation by the 26S proteasome, becomes phosphorylated in a cytokine-induced manner on three serines and one threonine. In this project three cytokine-activated kinases which phosphorylate FAT10 in vitro and in cells will be validated and further characterized. The consequences of FAT10 phosphorylation for its conjugation, for proteasomal degradation and for binding to p62 and HDAC6 and consecutive localization in aggresomes and autophagosomes will be investigated. The structures and conformational states of wild type, phosphomimetic, and phosphorylated FAT10 as well as two naturally occurring allelic variants will be investigated with magic-angle-spinning NMR and Molecular Dynamics Simulations, respectively. Moreover, patches on the surface of FAT10 required for binding to FAT10 ligands like the proteasome subunit Rpn10 and the linker protein Nub1L will be identified.

泛素的磷酸化对其在线粒体自噬中的功能具有重要影响。泛蛋白家族的另一种修饰剂，称为HLA-F邻近转录本10（FAT 10），也靶向缀合蛋白质以供26 S蛋白酶体降解，以细胞因子诱导的方式在三个丝氨酸和一个苏氨酸上磷酸化。在本项目中，将验证并进一步表征三种在体外和细胞中磷酸化FAT 10的精氨酸激活激酶。将研究FAT 10磷酸化对其缀合、蛋白酶体降解和与p62和HDAC 6结合以及在侵袭体和自噬体中连续定位的后果。野生型，磷酸化模拟，磷酸化脂肪10以及两个天然存在的等位基因变体的结构和构象状态将分别与魔角旋转NMR和分子动力学模拟进行研究。此外，将鉴定FAT 10表面上与FAT 10配体（如蛋白酶体亚基Rpn 10和接头蛋白Nub 1 L）结合所需的补丁。