Hubrecht Institute for Developmental Biology and Stem Cell Research, KNAW and University Medical Center Utrecht, Utrecht, The Netherlands.
Department of Nephrology and Hypertension, University Medical Center Utrecht, Utrecht, The Netherlands.
Oncogene. 2015 Feb 5;34(6):771-9. doi: 10.1038/onc.2013.598. Epub 2014 Jan 27.
Functional loss of the von Hippel-Lindau (VHL) tumor suppressor protein (pVHL), which is part of an E3-ubiquitin ligase complex, initiates most inherited and sporadic clear-cell renal cell carcinomas (ccRCC). Genetic inactivation of the TP53 gene in ccRCC is rare, suggesting that an alternate mechanism alleviates the selective pressure for TP53 mutations in ccRCC. Here we use a zebrafish model to describe the functional consequences of pVHL loss on the p53/Mdm2 pathway. We show that p53 is stabilized in the absence of pVHL and becomes hyperstabilized upon DNA damage, which we propose is because of a novel in vivo interaction revealed between human pVHL and a negative regulator of Mdm2, the programmed cell death 5 (PDCD5) protein. PDCD5 is normally localized at the plasma membrane and in the cytoplasm. However, upon hypoxia or loss of pVHL, PDCD5 relocalizes to the nucleus, an event that is coupled to the degradation of Mdm2. Despite the subsequent hyperstabilization and normal transcriptional activity of p53, we find that zebrafish vhl(-/-) cells are still as highly resistant to DNA damage-induced cell cycle arrest and apoptosis as human ccRCC cells. We suggest this is because of a marked increase in expression of birc5a, the zebrafish homolog of Survivin. Accordingly, when we knock down Survivin in human ccRCC cells we are able to restore caspase activity in response to DNA damage. Taken together, our study describes a new mechanism for p53 stabilization through PDCD5 upon hypoxia or pVHL loss, and reveals new clinical potential for the treatment of pathobiological disorders linked to hypoxic stress.
抑癌基因 von Hippel-Lindau(VHL)蛋白(pVHL)功能丧失,作为 E3 泛素连接酶复合物的一部分,引发大多数遗传性和散发性肾透明细胞癌(ccRCC)。ccRCC 中 TP53 基因的遗传失活很少见,这表明在 ccRCC 中存在另一种机制来缓解 TP53 突变的选择压力。在这里,我们使用斑马鱼模型来描述 pVHL 缺失对 p53/Mdm2 通路的功能影响。我们发现 p53 在没有 pVHL 的情况下稳定,并且在 DNA 损伤时变得更加稳定,我们提出这是因为在体内发现了人类 pVHL 和 Mdm2 的负调节剂程序性细胞死亡 5(PDCD5)蛋白之间的新相互作用。PDCD5 通常位于质膜和细胞质中。然而,在缺氧或 pVHL 缺失时,PDCD5 重新定位到细胞核,这与 Mdm2 的降解有关。尽管随后 p53 的稳定性增加且转录活性正常,但我们发现 vhl(-/-) 斑马鱼细胞对 DNA 损伤诱导的细胞周期阻滞和凋亡仍然高度耐受,与人类 ccRCC 细胞相似。我们认为这是因为 birc5a(Survivin 的斑马鱼同源物)的表达显著增加。因此,当我们在人 ccRCC 细胞中敲低 Survivin 时,我们能够恢复 caspase 活性以响应 DNA 损伤。总之,我们的研究描述了在缺氧或 pVHL 缺失时通过 PDCD5 稳定 p53 的新机制,并揭示了针对与缺氧应激相关的病理生物学紊乱的新的临床潜力。