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

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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.