Lang Valérie, Pallara Chiara, Zabala Amaia, Lobato-Gil Sofia, Lopitz-Otsoa Fernando, Farrás Rosa, Hjerpe Roland, Torres-Ramos Monica, Zabaleta Lorea, Blattner Christine, Hay Ronald T, Barrio Rosa, Carracedo Arkaitz, Fernandez-Recio Juan, Rodríguez Manuel S, Aillet Fabienne
Ubiquitylation and Cancer Molecular Biology Laboratory, Inbiomed, Mikeletegi 81, San Sebastián-Donostia 20009, Gipuzkoa, Spain.
Joint BSC-IRB Research Program in Computational Biology, Life Sciences Department, Barcelona Supercomputing Center, Carrer Jordi Girona 29, 08034 Barcelona, Spain.
Mol Oncol. 2014 Jul;8(5):1026-42. doi: 10.1016/j.molonc.2014.04.002. Epub 2014 Apr 13.
The tumor suppressor p53 regulates the expression of genes involved in cell cycle progression, senescence and apoptosis. Here, we investigated the effect of single point mutations in the oligomerization domain (OD) on tetramerization, transcription, ubiquitylation and stability of p53. As predicted by docking and molecular dynamics simulations, p53 OD mutants show functional defects on transcription, Mdm2-dependent ubiquitylation and 26S proteasome-mediated degradation. However, mutants unable to form tetramers are well degraded by the 20S proteasome. Unexpectedly, despite the lower structural stability compared to WT p53, p53 OD mutants form heterotetramers with WT p53 when expressed transiently or stably in cells wild type or null for p53. In consequence, p53 OD mutants interfere with the capacity of WT p53 tetramers to be properly ubiquitylated and result in changes of p53-dependent protein expression patterns, including the pro-apoptotic proteins Bax and PUMA under basal and adriamycin-induced conditions. Importantly, the patient derived p53 OD mutant L330R (OD1) showed the more severe changes in p53-dependent gene expression. Thus, in addition to the well-known effects on p53 stability, ubiquitylation defects promote changes in p53-dependent gene expression with implications on some of its functions.
肿瘤抑制因子p53可调节参与细胞周期进程、衰老和凋亡的基因表达。在此,我们研究了寡聚化结构域(OD)中的单点突变对p53四聚化、转录、泛素化和稳定性的影响。正如对接和分子动力学模拟所预测的,p53 OD突变体在转录、Mdm2依赖的泛素化和26S蛋白酶体介导的降解方面表现出功能缺陷。然而,无法形成四聚体的突变体可被20S蛋白酶体有效降解。出乎意料的是,尽管与野生型p53相比结构稳定性较低,但p53 OD突变体在野生型或p53缺失的细胞中瞬时或稳定表达时,可与野生型p53形成异源四聚体。因此,p53 OD突变体干扰了野生型p53四聚体被正确泛素化的能力,并导致p53依赖的蛋白质表达模式发生变化,包括在基础和阿霉素诱导条件下的促凋亡蛋白Bax和PUMA。重要的是,患者来源的p53 OD突变体L330R(OD1)在p53依赖的基因表达中表现出更严重的变化。因此,除了对p53稳定性的众所周知的影响外,泛素化缺陷还会促进p53依赖的基因表达变化,从而影响其某些功能。