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Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT

Rajan Kumar Choudhary1,2, M Quadir Siddiqui1,3, Nikhil Gadewal1

  • 1Advanced Centre for Treatment, Research and Education in Cancer Kharghar Navi Mumbai Maharashtra 410 210 India avarma@actrec.gov.in +91-22-2740 5085 +91-22-2740 5112.

RSC Advances
|May 13, 2022
PubMed

Insights

Mutations in the BARD1 BRCT domain affect its structural integrity and thermodynamic stability. Specifically, Val695Leu and Ser761Asn mutations significantly destabilize the protein, impacting its function in DNA damage repair.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Structural Biology

Background:

  • The BRCT domain of BARD1 (BARD1 BRCT) is crucial for DNA damage repair (DDR) and cell-cycle checkpoint regulation.
  • BARD1 BRCT acts as a tumor suppressor by facilitating BRCA1 recruitment to DNA damage sites.
  • Understanding the impact of BARD1 BRCT mutations is vital for genomic integrity.

Purpose of the Study:

  • To evaluate the pathogenicity of reported BARD1 BRCT mutations: Cys645Arg, Val695Leu, and Ser761Asn.
  • To investigate the effects of these mutations on BARD1 BRCT protein structure and function.
  • To correlate structural and functional changes with potential pathogenicity.

Main Methods:

  • Biochemical and biophysical techniques including fluorescence, circular dichroism (CD) spectroscopy, and limited proteolysis.
  • Thermal and chemical denaturation studies to assess protein stability.
  • In silico molecular dynamics (MD) simulations to analyze structural integrity.

Main Results:

  • Wild-type and mutant BARD1 BRCT proteins exhibit well-folded structural conformations.
  • Cys645Arg mutation shows folding patterns similar to wild-type BARD1 BRCT.
  • Val695Leu and Ser761Asn mutations lead to a significant loss in thermodynamic stability and structural integrity compared to wild-type.

Conclusions:

  • The Cys645Arg mutation may not significantly impair BARD1 BRCT's structural stability.
  • The Val695Leu and Ser761Asn mutations compromise the thermodynamic stability and structural integrity of BARD1 BRCT.
  • These findings highlight the functional consequences of specific BARD1 BRCT mutations on DNA damage repair pathways.

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