LZTR1 ve NUAK2 proteinlerinin etkileşimi ve mitokondriyal apoptoz yolağı üzerindeki etkileri

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Karabulut S., Yıldız G.

cilt.2, sa.3, ss.1-155, 2023 (Hakemli Dergi)

  • Yayın Türü: Makale / Özet
  • Cilt numarası: 2 Sayı: 3
  • Basım Tarihi: 2023
  • Derginin Tarandığı İndeksler: Index Copernicus
  • Sayfa Sayıları: ss.1-155
  • Açık Arşiv Koleksiyonu: AVESİS Açık Erişim Koleksiyonu
  • Karadeniz Teknik Üniversitesi Adresli: Evet

Özet

Objective: Leucine Zipper Like Post Translational Regulator 1 (LZTR1) has tumor suppressor functions in glioblastoma multiforme (GBM) and hepatocellular carcinoma (HCC), while NUAK Family Kinase 2 (NUAK2) has an oncogenic function. The aim of this study is to investigate the relationships between LZTR1 and NUAK2 and their effects on mitochondrial apoptosis pathway-induced cells.

Methods: For this purpose firstly, human embryonic kidney 293 (HEK293) cells stably expressing Flag-tagged LZTR1 were prepared. The interactions of LZTR1 and NUAK2 proteins and their intracellular localizations were examined by immunoprecipitation (IP) and immunofluorescence (IF) staining methods, respectively. Apoptosis was triggered by treating cells with 0.4 mM hydrogen peroxide (H2O2) for 10 h. BCL-2, BAX, p53, phosphorylated (p)-p53-Ser15, p-p53-Ser46, Survivin, Caspase 3, active Caspase 3, PARP1, and active PARP1 levels in parental HEK293 cells and stably LZTR1-expressing clones that were transiently transfected/ untransfected with NUAK2 expression plasmid or treated/non-treated with H2O2 were examined by western blotting (WB) method, using specific primary antibodies.

Results: IP experiments indicate that LZTR1 and NUAK2 interact. IF analyzes showed that LZTR1 and NUAK2 protein are found mostly in cytoplasm and nucles, respectively, alone; while in the presence of LZTR1, the localization of NUAK2 alters to be in the cytoplasm. Our WB results showed that active PARP1 protein levels increased with H2O2 treatment, LZTR1 levels decreased in apoptotic cells, LZTR1 caused decrease of NUAK2 levels, LZTR1 and NUAK2 co-expression in apoptotic cells caused decrease of Survivin but increase of p-p53-Ser46 levels.

Conclusion: With this study, interactions of LZTR1 and NUAK2 proteins, LZTR1-induced intracellular localization change of NUAK2, and the effects of LZTR1 and NUAK2 on H2O2-induced apoptosis were shown for the first time. These findings have potential to contribute to elucidation of the functions of LZTR1 and NUAK2 proteins and their roles in diseases, which they were associated with.

Keywords: LZTR1 and NUAK2 proteins, mitochondrial apoptosis pathway.
Acknowledgement: This work was supported by Office of Scientific Research Projects of Karadeniz Technical

University. Project number: TSA-2023-10539