Zhao Liuqun, Punga Tanel, Sanyal Suparna
Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Uppsala, Sweden.
Department of Medical Biochemistry and Microbiology, Uppsala University, Biomedical Center, Uppsala, Sweden.
Elife. 2025 Jul 21;14:RP106469. doi: 10.7554/eLife.106469.
p53 is a tumor suppressor protein with multiple isoforms with shared or specific functions. However, two of its isoforms, Δ133p53α and Δ160p53α, with large N-terminal deletions, can cause cancer. These isoforms exert a dominant-negative effect on full-length p53 (FLp53), although the precise molecular mechanisms are unknown. Here, we investigate the mechanisms of action of Δ133p53α and Δ160p53α isoforms using chromatin immunoprecipitation, luciferase expression, subcellular fractionation, immunofluorescence assays, and apoptotic caspase activity assay. Our study elucidates that these DNA-binding deficient p53 isoforms form hetero-tetrameric complexes with FLp53 and disrupt FLp53's DNA binding and transcriptional activities when present in a higher proportion than FLp53 in the tetramer. However, these structurally unstable isoforms promote vigorous protein aggregation involving FLp53, disrupting its structure and sequestering it in the cytoplasmic and nuclear aggregates, thereby limiting its availability to function as a transcription activator protein. Thus, co-aggregation of Δ133p53α and Δ160p53α with FLp53, rather than hetero-tetramerization, is likely the primary factor contributing to their dominant-negative effect. Modulating the stability and aggregation of p53 isoforms could be a novel strategy for cancer therapy.
p53是一种具有多种亚型的肿瘤抑制蛋白,这些亚型具有共同或特定的功能。然而,其两种亚型Δ133p53α和Δ160p53α,具有较大的N端缺失,可导致癌症。尽管确切的分子机制尚不清楚,但这些亚型对全长p53(FLp53)发挥显性负效应。在这里,我们使用染色质免疫沉淀、荧光素酶表达、亚细胞分级分离、免疫荧光测定和凋亡半胱天冬酶活性测定来研究Δ133p53α和Δ160p53α亚型的作用机制。我们的研究阐明,这些缺乏DNA结合能力的p53亚型与FLp53形成异源四聚体复合物,并且当在四聚体中以高于FLp53的比例存在时,会破坏FLp53的DNA结合和转录活性。然而,这些结构不稳定的亚型促进涉及FLp53的强烈蛋白质聚集,破坏其结构并将其隔离在细胞质和核聚集体中,从而限制其作为转录激活蛋白发挥功能的可用性。因此,Δ133p53α和Δ160p53α与FLp53的共聚集,而非异源四聚化,可能是导致其显性负效应的主要因素。调节p53亚型的稳定性和聚集可能是癌症治疗的一种新策略。