Isolation, characterization and functional analysis of full length p53 cDNA from Bubalus bubalis

Area of Science:

• Molecular Biology
• Genetics
• Cancer Research

Background:

• The p53 protein is crucial for genomic stability and tumor suppression.
• Understanding p53 function across species can reveal insights into its instability and therapeutic targeting.

Purpose of the Study:

• To characterize the full-length buffalo p53 cDNA (Bp53) and its functional properties.
• To compare Bp53's anti-tumorigenic activity and stability with human p53.

Main Methods:

• Cloning and expression of Bp53 cDNA in vectors for FLAG or GFP tagging.
• Transient transfection of Bp53 into a p53-null human lung cancer cell line.
• Assays for nuclear localization, gene transactivation, clonogenic survival, and cell viability.

Main Results:

• Bp53 shares high homology with human p53 but has distinct amino acid compositions.
• Bp53 localizes to the nucleus, transactivates target genes, and demonstrates tumor suppressor activity.
• A core domain mutant of Bp53 showed cytoplasmic mislocalization and reduced activity.
• Bp53 exhibited lower stability than human p53 due to heightened sensitivity to mdm2-mediated degradation.

Conclusions:

• Buffalo p53 is a functionally active wild-type protein with tumor suppressor capabilities.
• Bp53's reduced stability compared to human p53 offers a model for studying p53 instability.
• Further research on Bp53 could elucidate mechanisms underlying wild-type p53 instability.