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

赖氨酸-脱乙酰酶抑制剂、乙酰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能够鉴定急性髓性白血病（AML）细胞中所有细胞隔室中的数千种蛋白质为赖氨酸-乙酰化的。数据显示，细胞骨架似乎受到AML细胞中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，我将首先关注对几种特定蛋白质的研究（短期项目）。赖氨酸乙酰化通过hDia 1控制肌动蛋白成核和聚合，细胞内转运过程和小GNBP Ran的蛋白质周转，Cdc 42的效应子结合，通过RhoGEF 2的功能性Rho GTP/GDP循环，并最终协调代谢速率和细胞骨架组织以及糖酵解酶醛缩酶的酶活性。使用遗传密码扩展的概念，我们要纳入乙酰赖氨酸和分析乙酰化的影响在体外和研究蛋白质赖氨酸乙酰化的后果在细胞的基础上。此外，在中期，我们想了解乙酰化本身是如何调节的。这包括确定赖氨酸脱乙酰酶和乙酰转移酶的蛋白质，这是最初发现的功能受赖氨酸乙酰化。在长期的定量蛋白质组学中，使用SILAC将详细了解蛋白质的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的蛋白质活性和功能，这很可能支持衰老相关过程的发展，如肿瘤侵袭和转移。总之，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动，以及赖氨酸乙酰化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泛素化等翻译后修饰的功能差异，并可能开发新的治疗策略来治疗衰老相关疾病。

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.