Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Bengaluru 560064, India.
BioCOS Life Sciences Pvt. Ltd., Bengaluru, India.
J Biol Chem. 2019 Sep 20;294(38):14081-14095. doi: 10.1074/jbc.RA118.007265. Epub 2019 Jul 31.
TP53 is the most frequently mutated tumor suppressor gene in many cancers, yet biochemical characterization of several of its reported mutations with probable biological significance have not been accomplished enough. Specifically, missense mutations in TP53 can contribute to tumorigenesis through gain-of-function of biochemical and biological properties that stimulate tumor growth. Here, we identified a relatively rare mutation leading to a proline to leucine substitution (P152L) in TP53 at the very end of its DNA-binding domain (DBD) in a sample from an Indian oral cancer patient. Although the P152Lp53 DBD alone bound to DNA, the full-length protein completely lacked binding ability at its cognate DNA motifs. Interestingly, P152Lp53 could efficiently tetramerize, and the mutation had only a limited impact on the structure and stability of full-length p53. Significantly, when we expressed this variant in a TP53-null cell line, it induced cell motility, proliferation, and invasion compared with a vector-only control. Also, enhanced tumorigenic potential was observed when P152Lp53-expressing cells were xenografted into nude mice. Investigating the effects of P152Lp53 expression on cellular pathways, we found that it is associated with up-regulation of several pathways, including cell-cell and cell-extracellular matrix signaling, epidermal growth factor receptor signaling, and Rho-GTPase signaling, commonly active in tumorigenesis and metastasis. Taken together, our findings provide a detailed account of the biochemical and cellular alterations associated with the cancer-associated P152Lp53 variant and establish it as a gain-of-function TP53 variant.
TP53 是许多癌症中最常发生突变的肿瘤抑制基因,但对其一些报道的具有可能生物学意义的突变的生化特征研究还不够充分。具体来说,TP53 的错义突变可以通过刺激肿瘤生长的生化和生物学特性的功能获得来促进肿瘤发生。在这里,我们在一名印度口腔癌患者的样本中发现了一个相对罕见的突变,导致 DNA 结合域(DBD)末端的脯氨酸到亮氨酸取代(P152L)。尽管 P152Lp53 DBD 本身可以与 DNA 结合,但全长蛋白完全缺乏与其同源 DNA 基序的结合能力。有趣的是,P152Lp53 可以有效地四聚化,并且该突变对全长 p53 的结构和稳定性只有有限的影响。重要的是,当我们在 TP53 缺失的细胞系中表达这种变体时,与仅载体对照相比,它会诱导细胞迁移、增殖和侵袭。此外,当 P152Lp53 表达细胞被异种移植到裸鼠中时,观察到增强的致瘤潜力。研究 P152Lp53 表达对细胞通路的影响,我们发现它与包括细胞-细胞和细胞-细胞外基质信号、表皮生长因子受体信号和 Rho-GTPase 信号在内的几条通路的上调有关,这些通路在肿瘤发生和转移中通常是活跃的。总之,我们的研究结果提供了与癌症相关的 P152Lp53 变体相关的生化和细胞改变的详细描述,并将其确立为一种具有功能获得的 TP53 变体。