Attenuation of DNA damage checkpoint by PBK, a novel mitotic kinase, involves protein–protein interaction with tumor suppressor p53
Section snippets
Materials and methods
Plasmids and cell line. HT 1080 fibrosarcoma cell line was obtained from American Type Culture Collection, Manassas, VA (#CCL-121). Yeast vectors pGBKT7 and pGADT7 were utilized for expressing PBK and p53 as fusion proteins with Gal4 DNA-binding domain and activation domain, respectively.
Western immunoblotting and immunoprecipitations. Western immunoblotting of cell lysates derived from PBK expressing clones were carried out as described previously [25]. Monoclonal PBK antibody and
Results
Upon examination by immunoblot analysis, fibrosarcoma cells (HT1080) showed low levels of expression of endogenous PBK gene as compared to HL60 promyelocytic leukemia cell line (Fig. 1A). However, PBK expression was negatively regulated during growth arrest predominantly in the G2/M phase by the presence of low doses of an antineoplastic compound doxorubicin, similar to leukemia cells (Fig. 1B) [30]. Thus in the presence of doxorubicin, PBK protein level diminished slowly until it was barely
Discussion
Identifying the cellular machinery which attributes to genomic instabilities of various kinds is crucial for understanding the mechanisms accumulating further genetic alterations that lead to malignant diseases. The genes that are pertinent to this function could be worthwhile targets for cancer therapeutics and are also of immense prognostic value. Logically, emphasis are being bestowed upon a number of cell cycle checkpoints which provide surveillance towards maintaining fidelity at multiple
Acknowledgments
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2023, GeneCitation Excerpt :Although, the PBK expression promoted migration and invasion of GC cell lines (such as SNU638 and AGS); however, its expression was not sufficient for cell proliferation (Kwon, 2016). PBK reduces the expression of P53 and P38-MAPK, which in turn suppresses tumor cell mortality (Ayllon and O'connor, 2007; Nandi, 2007; Hu, 2010). The interaction of PBK with the PI3K/PTEN/AKT axis promoted cell migration, introducing the oncogenic role of PBK via P53 and the PI3K/AKT pathways in GC (Ohashi, 2017).
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2021, Molecular and Cellular ProteomicsCitation Excerpt :PBK/TOPK (PDZ-binding kinase/T-LAK cell-originated protein kinase) is involved in control of the cell cycle and mitotic progression (42). In fibrosarcoma cells it was shown that, upon doxorubicin-induced DNA damage and PBK overexpression, cells bypassed the G2/M DNA damage checkpoint and entered into the next mitosis (43), which might explain the need of cells to regulate PBK activity during situations of DNA damage. Taken together, our comparison of untreated and irradiated U2OS cells reproduces previous findings on kinase activity changes upon induction of DNA damage and identified kinase activity changes are largely overlapping for both extraction methods.
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2019, European Journal of Medicinal ChemistryCitation Excerpt :Also, studies have shown that TOPK could phosphorylate histone H3 at Ser10 and serves as a molecular marker in breast cancer [3]. Increased levels of PBK/TOPK may contribute to tumor cell development and progression through suppression of p53 function [13,16]. TOPK was also reported to increase cell migration by modulating a PI3K/PTEN/AKT-dependent signaling pathway [17].