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.

赖氨酸去乙酰化酶抑制剂、赖氨酸特异性抗体的可用性以及定量蛋白质组学的巨大进展，最近使得在急性髓性白血病（AML）细胞中，所有细胞区室中数千种蛋白质被赖氨酸乙酰化。数据显示，AML细胞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。为了证明乙酰化可以发挥的多种调节作用，我将首先关注几种特定蛋白质的研究（短期项目）。赖氨酸乙酰化通过hDia1控制肌动蛋白成核和聚合，细胞内运输过程和小GNBP Ran的蛋白质周转，Cdc42的效应结合，通过RhoGEF2的Rho GTP/GDP循环，并最终协调代谢率和细胞骨架组织以及糖酵解酶醛缩酶的酶活性。利用遗传密码扩展的概念，我们希望纳入乙酰赖氨酸，并分析乙酰化在体外的作用，并研究蛋白质赖氨酸乙酰化在细胞基础上的后果。此外，在中期，我们希望了解乙酰化本身是如何调节的。这包括鉴定蛋白质的赖氨酸去乙酰化酶和-乙酰转移酶，它们最初被发现在功能上受到赖氨酸乙酰化的影响。在长期的定量蛋白质组学中，使用SILAC将提供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如何变化的详细视图。如果细胞骨架乙酰化模式的改变与蛋白质活性和功能的改变相对应，这很可能支持衰老相关过程的发展，如肿瘤侵袭和转移。综上所述，我们想了解为什么这么多参与细胞骨架调节的蛋白质被乙酰化，乙酰化如何控制这些活性，以及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。这些研究将揭示赖氨酸乙酰化与磷酸化和泛素化等翻译后修饰的功能差异，并可能开发出治疗衰老相关疾病的新治疗策略。