Ghosh Ginia, Misra Sandip, Ray Rachayeeta, Chowdhury Sougata Ghosh, Karmakar Parimal
Department of Life Science and Biotechnology, Jadavpur University, Kolkata, West Bengal, India.
Department of Microbiology, Bidhannagar College, Salt Lake, Kolkata, West Bengal, India.
Med Oncol. 2023 Mar 17;40(4):119. doi: 10.1007/s12032-023-01985-z.
PTEN, dual phosphatase tumor suppressor protein, is found to be frequently mutated in various cancers. Post-translational modification of PTEN is important for its sub-cellular localization and catalytic functions. But how these modifications affect cytological damage and aneuploidy is not studied in detail. We focus on the role of phosphatase activity along with C-terminal phosphorylation of PTEN in perspective of cytological damage like micronucleus, nuclear bud, and nuclear bridge formation. Our data suggest that wild-type PTEN, but not phospho-mutant PTEN significantly reduces cytological damage in PTEN null PC3 cells. In case of phosphatase-dead PTEN, cytological damage markers are increased during 24 h recovery after DNA damage. When we use phosphorylation and phosphatase-dead dual mutant PTEN, the extent of different cytological DNA damage parameters are similar to phosphatase-dead PTEN. We also find that both of those activities are essential for maintaining chromosome numbers. PTEN null cells exhibit significantly aberrant γ-tubulin pole formation during metaphase. Interestingly, we observed that p-PTEN localized to spindle poles along with PLK1 and Aurora Kinase A. Further depletion of phosphorylation and phosphatase activity of PTEN increases the expression of p-Aurora Kinase A (T288) and p-PLK1 (T210), compared to cells expressing wild-type PTEN. Again, wild-type PTEN but not phosphorylation-dead mutant is able to physically interact with PLK1 and Aurora Kinase A. Thus, our study suggests that the phosphorylation-dependent interaction of PTEN with PLK1 and Aurora Kinase A causes dephosphorylation of those mitotic kinases and by lowering their hyperphosphorylation status, PTEN prevents aberrant chromosome segregation in metaphase.
磷酸酶与张力蛋白同源物(PTEN)是一种双重磷酸酶肿瘤抑制蛋白,在多种癌症中经常发生突变。PTEN的翻译后修饰对其亚细胞定位和催化功能很重要。但这些修饰如何影响细胞损伤和非整倍体尚未得到详细研究。我们从微核、核芽和核桥形成等细胞损伤角度,重点研究了PTEN磷酸酶活性及其C末端磷酸化的作用。我们的数据表明,野生型PTEN而非磷酸化突变型PTEN能显著降低PTEN基因缺失的PC3细胞中的细胞损伤。在磷酸酶失活的PTEN情况下,DNA损伤后24小时恢复期间细胞损伤标志物增加。当我们使用磷酸化和磷酸酶失活的双重突变型PTEN时,不同细胞DNA损伤参数的程度与磷酸酶失活的PTEN相似。我们还发现这两种活性对于维持染色体数量至关重要。PTEN基因缺失的细胞在中期表现出明显异常的γ-微管蛋白极形成。有趣的是,我们观察到磷酸化的PTEN与PLK1和极光激酶A一起定位于纺锤体极。与表达野生型PTEN的细胞相比,进一步耗尽PTEN的磷酸化和磷酸酶活性会增加磷酸化的极光激酶A(T288)和磷酸化的PLK1(T210)的表达。同样,野生型PTEN而非磷酸化失活突变体能与PLK1和极光激酶A发生物理相互作用。因此,我们的研究表明,PTEN与PLK1和极光激酶A的磷酸化依赖性相互作用导致这些有丝分裂激酶去磷酸化,并且通过降低它们的过度磷酸化状态,PTEN可防止中期异常染色体分离。