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

The BRCT domain of BARD1 (BARD1 BRCT) is involved in many cellular processes such as DNA damage repair (DDR) and cell-cycle checkpoint regulation. BARD1 BRCT performs tumor suppressor function by recruiting BRCA1 at DNA damage site via interactions with other DNA damage repair (DDR) proteins. Consid...

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Autores principales: Choudhary, Rajan Kumar, Siddiqui, M. Quadir, Gadewal, Nikhil, Kumar, Nachimuthu Senthil, Kuligina, Ekaterina S., Varma, Ashok K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9086705/
https://www.ncbi.nlm.nih.gov/pubmed/35548793
http://dx.doi.org/10.1039/c8ra06524a
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author Choudhary, Rajan Kumar
Siddiqui, M. Quadir
Gadewal, Nikhil
Kumar, Nachimuthu Senthil
Kuligina, Ekaterina S.
Varma, Ashok K.
author_facet Choudhary, Rajan Kumar
Siddiqui, M. Quadir
Gadewal, Nikhil
Kumar, Nachimuthu Senthil
Kuligina, Ekaterina S.
Varma, Ashok K.
author_sort Choudhary, Rajan Kumar
collection PubMed
description The BRCT domain of BARD1 (BARD1 BRCT) is involved in many cellular processes such as DNA damage repair (DDR) and cell-cycle checkpoint regulation. BARD1 BRCT performs tumor suppressor function by recruiting BRCA1 at DNA damage site via interactions with other DNA damage repair (DDR) proteins. Considering the importance of the BRCT domain in genomic integrity, we decided to evaluate reported mutations of BARD1 BRCT Cys645Arg, Val695Leu, and Ser761Asn for their pathogenicity. To explore the effect of the mutation on the structure and function, BARD1 BRCT wild-type proteins and the mutant proteins were studied using different biochemical, biophysical and in silico techniques. Comparative fluorescence, circular dichroism (CD) spectroscopy and limited proteolysis studies demonstrate the well-folded structural conformation of wild-type and mutant proteins. However, thermal and chemical denaturation studies revealed similarity in the folding pattern of BARD1 BRCT wild-type and Cys645Arg mutant proteins, whereas there was a significant loss in the thermodynamic stability of Val695Leu and Ser761Asn mutants. Molecular dynamics (MD) simulation studies on wild-type and mutant protein structures indicate the loss in structural integrity of mutants compared with the wild-type protein.
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spelling pubmed-90867052022-05-10 Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT domain Choudhary, Rajan Kumar Siddiqui, M. Quadir Gadewal, Nikhil Kumar, Nachimuthu Senthil Kuligina, Ekaterina S. Varma, Ashok K. RSC Adv Chemistry The BRCT domain of BARD1 (BARD1 BRCT) is involved in many cellular processes such as DNA damage repair (DDR) and cell-cycle checkpoint regulation. BARD1 BRCT performs tumor suppressor function by recruiting BRCA1 at DNA damage site via interactions with other DNA damage repair (DDR) proteins. Considering the importance of the BRCT domain in genomic integrity, we decided to evaluate reported mutations of BARD1 BRCT Cys645Arg, Val695Leu, and Ser761Asn for their pathogenicity. To explore the effect of the mutation on the structure and function, BARD1 BRCT wild-type proteins and the mutant proteins were studied using different biochemical, biophysical and in silico techniques. Comparative fluorescence, circular dichroism (CD) spectroscopy and limited proteolysis studies demonstrate the well-folded structural conformation of wild-type and mutant proteins. However, thermal and chemical denaturation studies revealed similarity in the folding pattern of BARD1 BRCT wild-type and Cys645Arg mutant proteins, whereas there was a significant loss in the thermodynamic stability of Val695Leu and Ser761Asn mutants. Molecular dynamics (MD) simulation studies on wild-type and mutant protein structures indicate the loss in structural integrity of mutants compared with the wild-type protein. The Royal Society of Chemistry 2018-10-03 /pmc/articles/PMC9086705/ /pubmed/35548793 http://dx.doi.org/10.1039/c8ra06524a Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Choudhary, Rajan Kumar
Siddiqui, M. Quadir
Gadewal, Nikhil
Kumar, Nachimuthu Senthil
Kuligina, Ekaterina S.
Varma, Ashok K.
Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT domain
title Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT domain
title_full Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT domain
title_fullStr Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT domain
title_full_unstemmed Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT domain
title_short Biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the BARD1 BRCT domain
title_sort biophysical evaluation to categorize pathogenicity of cancer-predisposing mutations identified in the bard1 brct domain
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9086705/
https://www.ncbi.nlm.nih.gov/pubmed/35548793
http://dx.doi.org/10.1039/c8ra06524a
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