Untersuchung des Einflusses post-translationaler Lysin Acetylierung als globaler Regulator des Zytoskeletts

研究翻译后赖氨酸乙酰化作为细胞骨架全局调节剂的影响

• 批准号: 200568056
• 负责人: Professor Dr. Michael Lammers
• 金额: --
• 依托单位: Institut für Biochemie
• 依托单位国家: 德国
• 项目类别: Independent Junior Research Groups
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/200568056?language=en
• 关键词: Untersuchung; des; Einflusses; post; translationaler

The availability of lysine-deacetylase inhibitors, acetyl-lysine specific antibodies and the enormous progress in quantitative proteomics recently enabled the identification of thousands of proteins in all cellular compartments as being lysine-acetylated in acute myeloid leukemia (AML) cells. The data revealed that the cytoskeleton appears to be highly affected by protein acetylation in AML cells, but how acetylation regulates protein-function remained unclear. To demonstrate the diverse regulative roles acetylation can perform, I will focus initially on studies on several specific proteins (short-term projects). Lysine-acetylation controls actin-nucleation and -polymerisation via hDia1, intracellular transport processes and protein-turnover of the small GNBP Ran, effector-binding of Cdc42, the functional Rho GTP/GDP cycle via RhoGEF2 and finally coordinates metabolic rates and cytoskeletal organisation as well as enzymatic activity of the glycolytic enzyme aldolase. Using the genetic-code expansion concept we want to incorporate acetyl-lysine and analyse the effect of acetylation in vitro and to study the consequences of protein lysine-acetylation on the cellular basis. Furthermore, in the mid-term we want to understand how acetylation itself is regulated. This includes identification of lysine deacetylases and – acetyltransferases for proteins, which were initially found to be functionally affected by lysine-acetylation. In the long-term quantitative proteomics using SILAC will give a detailed view of how the acetylation pattern of proteins change in ageing-associated dieseases as cancer and neurodegenerative disorders. If the change in the cytoskeletal acetylation pattern corresponds to altered protein activities and functionalities, this very likely supports development of ageing-associated processes as tumor-invasion and -metastasis. As a summary, we want to understand why so many proteins involved in cytoskeleton regulation are acetylated, how acetylation controls those activities and finally how the dysfunction of lysine-acetylation is a prerequisite for ageing-associated disorders. These studies will show the functional differences of lysine-acetylation compared to post-translational modifications as phosphorylation and ubiquitylation and may enable the development of new therapeutic strategies to treat ageing-associated dieseases.

赖氨酸脱乙酰酶抑制剂、乙酰赖氨酸特异性抗体的出现以及定量蛋白质组学的巨大进展最近使得能够在急性髓性白血病 (AML) 细胞中鉴定出所有细胞区室中的数千种蛋白质被赖氨酸乙酰化。 AML 细胞中的细胞骨架似乎受到蛋白质乙酰化的高度影响，但乙酰化如何调节蛋白质功能仍不清楚。由于乙酰化可以发挥常规作用，我首先将重点研究几种特定蛋白质（短期项目），赖氨酸乙酰化通过 hDia1 控制肌动蛋白成核和聚合、细胞内转运过程和小 GNBP Ran 效应器的蛋白质周转。 -通过 RhoGEF2 结合 Cdc42（功能性 Rho GTP/GDP 循环），最终协调代谢率和细胞骨架组织以及酶活性使用遗传密码扩展概念，我们希望合并乙酰赖氨酸并分析体外乙酰化的影响，并研究蛋白质赖氨酸乙酰化对细胞基础的影响。想要了解乙酰化本身是如何调节的，这包括蛋白质的赖氨酸脱乙酰酶和乙酰转移酶的鉴定，这些酶最初被发现具有功能性。在长期定量蛋白质组学中，使用 SILAC 可以详细了解蛋白质乙酰化模式在癌症和神经退行性疾病等衰老相关疾病中的变化（如果细胞骨架乙酰化模式的变化对应于改变）。蛋白质的活性和功能，这很可能支持肿瘤侵袭和转移等与衰老相关的过程的发展。总而言之，我们想了解为什么有这么多蛋白质。参与细胞骨架调节的氨基酸被乙酰化，乙酰化如何控制这些活性，最后赖氨酸乙酰化的功能障碍如何成为衰老相关疾病的先决条件。这些研究将显示赖氨酸乙酰化与磷酸化和翻译后修饰相比的功能差异。泛素化并可能有助于开发新的治疗策略来治疗与衰老相关的疾病。