title: Multimodal approach to explore the pathogenicity of BARD1, ARG 658 CYS, and ILE 738 VAL mutants.
abstract: BARD1-BRCA1 complex plays an important role in DNA damage repair, apoptosis, chromatin remodeling, and other important processes required for cell survival. BRCA1 and BARD1 heterodimer possess E3 ligase activity and is involved in genome maintenance, by functioning in surveillance for DNA damage, thereby regulating multiple pathways including tumor suppression. BRCT domains are evolutionary conserved domains present in different proteins such as BRCA1, BARD1, XRCC, and MDC1 regulating damage response and cell-cycle control through protein-protein interactions. Nonetheless, the role of BARD1BRCT in the recruitment of DNA repair mechanism and structural integrity with BRCA1 complex is still implicit. To explicate the role of BARD1BRCT in the DNA repair mechanism, in silico, in vitro, and biophysical approach were applied to characterize BARD1 BRCT wild-type and Arg658Cys and Ile738Val mutants. However, no drastic secondary and tertiary structural changes in the mutant proteins were observed. Thermal and chemical denaturation studies revealed that mutants Arg658Cys and Ile738Val have a decrease in Tm and ∆G than the wild type. In silico studies of BARD1 BRCT (568-777) and mutant protein indicate loss in structural compactness on the Ile738Val mutant. Comparative studies of wild-type and mutants will thus be helpful in understanding the basic role of BARD1BRCT in DNA damage repair.
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: Rajan Kumar
authors: RK
authors: Choudhary
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: Vikrant
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: Quadir M
authors: QM
authors: Siddiqui
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: Pankaj S
authors: PS
authors: Thapa
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: Sweta
authors: S
authors: Raikundalia
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: Nikhil
authors: N
authors: Gadewal
authors: b Department of Biotechnology , Mizoram University (A Central University) , Aizawl 796 004 , Mizoram , India.
authors: Nachimuthu Senthil
authors: NS
authors: Kumar
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: M V
authors: MV
authors: Hosur
authors: a Advanced Centre for Treatment, Research and Education in Cancer , Kharghar , Navi Mumbai 410 210 , Maharashtra , India.
authors: Ashok K
authors: AK
authors: Varma
mesh_terms: D019943:Amino Acid Substitution; D003062:Codon; D006801:Humans; D008958:Models, Molecular; D050505:Mutant Proteins; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D055550:Protein Stability; D013057:Spectrum Analysis; D013329:Structure-Activity Relationship; D013816:Thermodynamics; D025521:Tumor Suppressor Proteins / Q000737:chemistry* / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry* / Q000378:metabolism
chemical_list: D003062:Codon; D050505:Mutant Proteins; D025521:Tumor Suppressor Proteins; C102449:BARD1 protein, human; D044767:Ubiquitin-Protein Ligases
keywords: BARD1 BRCT; protein–protein interactions; secondary and tertiary structure; thermal and chemical denaturation