Research

Life Sciences & Biotechnology

Title :

Unraveling p53 proteins amyloid aggregation and Liquid-Liquid Phase Separation dynamics in cancer

Area of research :

Life Sciences & Biotechnology

Principal Investigator :

Dr. Swathi Sudhakar, Indian Institute Of Technology (IIT) Madras, Tamil Nadu

Timeline Start Year :

2024

Timeline End Year :

2026

Contact info :

Equipments :

Details

Executive Summary :

The anti-oncogenic protein p53 plays a crucial role in cellular functions such as cell division, DNA repair, and apoptosis. However, its anti-oncogenic function is lost in over 50% of human cancer due to a mutated DNA-binding domain that is highly prone to prion-like aggregation. The molecular mechanisms of p53 amyloid aggregation behavior in cancer remain unclear, and real-time elucidation of this behavior is needed to understand their loss of function. The protein system's multivalent macromolecular interactions, intrinsically disordered regions, and patterned intermolecular electrostatic interactions drive Liquid Liquid phase separation (LLPS). In Alzheimer's, amyloid beta, a prion-like protein similar to p53, forms liquid-liquid phase-separated droplets enhanced with fibrils. However, the relationship between p53 amyloid aggregation and LLPS remains unclear. Polyphosphate (PP), a polyanionic inorganic polymer, is highly associated with cancer cell proliferation and progression. Studies suggest that PP accelerates fibril formation and LLPS in IDP, but its role in p53 amyloid aggregation and LLPS and their relation to cancer progression is yet to be understood. The proposed research aims to elucidate the prion-like fibril-forming mechanism of mutated and wild-type p53 by reconstituting the assay in flow cells and visualizing their fibril formation in real time using Total Internal Reflection Microscopy (TIRFM). The secondary structural conformation of the p53 protein associated with mutated and wild-type p53 will be studied using ThT fluorescence assay and CD spectroscopic techniques. This study will provide insights into p53 amyloidosis and LLPS mechanism, and pave the way for next-generation targeted cancer therapy.

Total Budget (INR):

32,97,310

Organizations involved