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Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
Wild type p53 function in p53Y220C mutant harboring cells by treatment with Ashwagandha derived anticancer
Durai Sundar1, Yue Yu2, Shashank P Katiyar1
1DAILAB, Department of Biochemical Engineering & Biotechnology, Indian Institute of Technology (IIT) Delhi, Hauz Khas, New Delhi, 110 016, India.
Background:
Tumor suppressor p53 protein is frequently mutated in a large majority of cancers. These mutations induce local or global changes in protein structure thereby affecting its binding to DNA. The structural differences between the wild type and mutant p53 thus provide an opportunity to selectively target mutated p53 harboring cancer cells. Restoration of wild type p53 activity in mutants using small molecules that can revert the structural changes have been considered for cancer therapeutics.
Methods:
We used bioinformatics and molecular docking tools to investigate the structural changes between the wild type and mutant p53 proteins (p53V143A, p53R249S, p53R273H and p53Y220C) and explored the therapeutic potential of Withaferin A and Withanone for restoration of wild type p53 function in cancer cells. Cancer cells harboring the specific mutant p53 proteins were used for molecular assays to determine the mutant or wild type p53 functions.
Results:
We found that p53V143A mutation does not show any significant structural changes and was also refractory to the binding of withanolides. p53R249S mutation critically disturbed the H-bond network and destabilized the DNA binding site. However, withanolides did not show any selective binding to either this mutant or other similar variants. p53Y220C mutation created a cavity near the site of mutation with local loss of hydrophobicity and water network, leading to functionally inactive conformation. Mutated structure could accommodate withanolides suggesting their conformational selectivity to target p53Y220C mutant. Using human cell lines containing specific p53 mutant proteins, we demonstrated that Withaferin A, Withanone and the extract rich in these withanolides caused restoration of wild type p53 function in mutant p53Y220C cells. This was associated with induction of p21WAF-1-mediated growth arrest/apoptosis.
Conclusion:
The study suggested that withanolides may serve as highly potent anticancer compounds for treatment of cancers harboring a p53Y220C mutation.
Insights
Withanolides, like Withaferin A and Withanone, show potential in restoring wild type p53 protein function in cancers with the specific p53Y220C mutation, offering a new therapeutic avenue.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Tumor suppressor p53 protein is frequently mutated in many cancers, altering its DNA binding and function.
- Mutations in p53 lead to structural changes, creating opportunities for targeted therapies.
- Restoring wild type p53 activity in cancer cells is a therapeutic strategy.
Purpose of the Study:
- To investigate structural differences between wild type and mutant p53 proteins.
- To explore the therapeutic potential of Withaferin A and Withanone for restoring wild type p53 function.
- To identify specific p53 mutations amenable to small molecule intervention.
Main Methods:
- Bioinformatics and molecular docking were used to analyze p53 protein structures.
- Investigated structural changes in p53V143A, p53R249S, p53R273H, and p53Y220C mutants.
- Utilized molecular assays in cancer cell lines harboring specific p53 mutations.
Main Results:
- p53V143A mutations showed no significant structural changes and were unresponsive to withanolides.
- p53R249S mutations destabilized the DNA binding site, but withanolides did not selectively bind.
- p53Y220C mutations created a cavity accommodating withanolides, leading to restoration of wild type p53 function and apoptosis induction.
- Withaferin A and Withanone restored wild type p53 function in p53Y220C mutant cells.
Conclusions:
- Withanolides demonstrate conformational selectivity for the p53Y220C mutant.
- Withaferin A and Withanone show promise as anticancer agents for p53Y220C-mutated cancers.
- These findings suggest a potential therapeutic strategy targeting specific p53 mutations.
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