期刊
CHEMICAL SCIENCE
卷 12, 期 21, 页码 7334-7349出版社
ROYAL SOC CHEMISTRY
DOI: 10.1039/d1sc01739j
关键词
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资金
- Carlos Chagas Filho Foundation for Research Support in the State of Rio de Janeiro (FAPERJ) [210.008/2018, 202840/2018]
- National Council for Scientific and Technological Development (CNPq)
- National Institute of Science and Technology for Structural Biology and Bioimaging (INCT) [465395/2014-7, 402321/2016-2]
- Pew Charitable Trusts Foundation
Mutant p53 tends to form aggregates with amyloid properties, especially amyloid oligomers inside the nucleus, which are believed to cause oncogenic gain-of-function. The DNA-binding domain of p53 (p53C) undergoes phase separation and aggregation in the presence of polyethylene glycol (PEG), and mutant p53C undergoes phase separation faster than wild-type p53C. Mutant p53 undergoes gel- and solid-like phase transitions in the nucleus, modulated by polyanions like heparin and RNA. The disordered transactivation domain (TAD) also plays a role in phase separation and amyloid aggregation of p53.
Mutant p53 tends to form aggregates with amyloid properties, especially amyloid oligomers inside the nucleus, which are believed to cause oncogenic gain-of-function (GoF). The mechanism of the formation of the aggregates in the nucleus remains uncertain. The present study demonstrated that the DNA-binding domain of p53 (p53C) underwent phase separation (PS) on the pathway to aggregation under various conditions. p53C phase separated in the presence of the crowding agent polyethylene glycol (PEG). Similarly, mutant p53C (M237I and R249S) underwent PS; however, the process evolved to a solid-like phase transition faster than that in the case of wild-type p53C. The data obtained by microscopy of live cells indicated that transfection of mutant full-length p53 into the cells tended to result in PS and phase transition (PT) in the nuclear compartments, which are likely the cause of the GoF effects. Fluorescence recovery after photobleaching (FRAP) experiments revealed liquid characteristics of the condensates in the nucleus. Mutant p53 tended to undergo gel- and solid-like phase transitions in the nucleus and in nuclear bodies demonstrated by slow and incomplete recovery of fluorescence after photobleaching. Polyanions, such as heparin and RNA, were able to modulate PS and PT in vitro. Heparin apparently stabilized the condensates in a gel-like state, and RNA apparently induced a solid-like state of the protein even in the absence of PEG. Conditions that destabilize p53C into a molten globule conformation also produced liquid droplets in the absence of crowding. The disordered transactivation domain (TAD) modulated both phase separation and amyloid aggregation. In summary, our data provide mechanistic insight into the formation of p53 condensates and conditions that may result in the formation of aggregated structures, such as mutant amyloid oligomers, in cancer. The pathway of mutant p53 from liquid droplets to gel-like and solid-like (amyloid) species may be a suitable target for anticancer therapy.
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