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Functional evaluation of gr acetylation in ahepatocyte cell line

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  1. Tez No: 786791
  2. Yazar: ULAŞ KAPLAN
  3. Danışmanlar: PROF. DR. THORSTEN HEİNZEL, PROF. DR. ARİA BANİAHMAD
  4. Tez Türü: Yüksek Lisans
  5. Konular: Biyokimya, Biochemistry
  6. Anahtar Kelimeler: Belirtilmemiş.
  7. Yıl: 2019
  8. Dil: İngilizce
  9. Üniversite: Friedrich-Schiller-Universität Jena
  10. Enstitü: Yurtdışı Enstitü
  11. Ana Bilim Dalı: Belirtilmemiş.
  12. Bilim Dalı: Belirtilmemiş.
  13. Sayfa Sayısı: 71

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Özet (Çeviri)

Glucocorticoids (GCs) are hormones that are named after their function in metabolism. However, their function is not limited to metabolism but extending to the immune system, central nervous system, musculoskeletal system, and cardiovascular system to name a few. To exert their function, GCs bind to their receptor called glucocorticoid receptor (GR) in the cytoplasm. After the binding of the GCs, GRs are activated and localized in the nucleus. Over there, GRs regulate expression of different genes through different mechanisms to regulate the variety of functions that are mentioned. One of these mechanisms is thought to be related to the epigenetic changes. The acetylation of GR is one of the less known epigenetic modifications of GR. It has been claimed that the hinge region of GR can be acetylated. Two different studies, one of which focused on the lysine acetyltransferase and the other focused on the lysine deacetylase have shown different functions of GR acetylation. Then, a study in our lab has revealed seven lysine residues in the N-terminal region of GR can be acetylated. It has also been shown that SIRT1 is the main deacetylase for the GR acetylation. Lysine to arginine mutations of the GR acetylation sites with site-directed mutagenesis illustrated that the K154 site could be important as the mutation of K154 abolished overall acetylation of GR. Moreover, a recent publication argues that SIRT1 increases the GR target gene expression without deacetylase activity on GR. The aim of this study is to check the function of GR acetylation in a physiologically relevant cell system. For this purpose, mainly Hepa1-6 cells were used. These cells are mouse hepatoma cells and with this study acetylation of the homologous site of GR K154 in mice was confirmed. Consequently, Sirt1 was found to be a deacetylase for this site and it was hinted that other sirtuins may deacetylate the same site as well. In addition to Hepa1-6 cells, macrophages were used to check GR K154 acetylation and its deacetylase as well. In macrophages, the acetylation of this particular site was deduced and further studies are needed for the function of Sirt1 on this site in macrophages. Then, qPCR experiments with Sirt1 and Sirt2 inhibitors showed that the inhibition of Sirt1 can increase, decrease or doesn't change the GR target gene expressions depending on the genes. For further experiments, the direct effect of GR acetylation and GR K154 acetylation should be established

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