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Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico

BACKGROUND: DNA topoisomerases are key enzymes that modulate the topological state of DNA through the breaking and rejoining of DNA strands. Human topoisomerase IB can be inhibited by several compounds that act through different mechanisms, including clinically used drugs, such as the derivatives of...

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Autores principales: Tesauro, Cinzia, Morozzo della Rocca, Blasco, Ottaviani, Alessio, Coletta, Andrea, Zuccaro, Laura, Arnò, Barbara, D'Annessa, Ilda, Fiorani, Paola, Desideri, Alessandro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3766703/
https://www.ncbi.nlm.nih.gov/pubmed/24004603
http://dx.doi.org/10.1186/1476-4598-12-100
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author Tesauro, Cinzia
Morozzo della Rocca, Blasco
Ottaviani, Alessio
Coletta, Andrea
Zuccaro, Laura
Arnò, Barbara
D'Annessa, Ilda
Fiorani, Paola
Desideri, Alessandro
author_facet Tesauro, Cinzia
Morozzo della Rocca, Blasco
Ottaviani, Alessio
Coletta, Andrea
Zuccaro, Laura
Arnò, Barbara
D'Annessa, Ilda
Fiorani, Paola
Desideri, Alessandro
author_sort Tesauro, Cinzia
collection PubMed
description BACKGROUND: DNA topoisomerases are key enzymes that modulate the topological state of DNA through the breaking and rejoining of DNA strands. Human topoisomerase IB can be inhibited by several compounds that act through different mechanisms, including clinically used drugs, such as the derivatives of the natural compound camptothecin that reversibly bind the covalent topoisomerase-DNA complex, slowing down the religation of the cleaved DNA strand, thus inducing cell death. Three enzyme mutations, which confer resistance to irinotecan in an adenocarcinoma cell line, were recently identified but the molecular mechanism of resistance was unclear. METHODS: The three resistant mutants have been investigated in S. cerevisiae model system following their viability in presence of increasing amounts of camptothecin. A systematical analysis of the different catalytic steps has been made for one of these mutants (Glu710Gly) and has been correlated with its structural-dynamical properties studied by classical molecular dynamics simulation. RESULTS: The three mutants display a different degree of camptothecin resistance in a yeast cell viability assay. Characterization of the different steps of the catalytic cycle of the Glu710Gly mutant indicated that its resistance is related to a high religation rate that is hardly affected by the presence of the drug. Analysis of the dynamic properties through simulation indicate that the mutant displays a much lower degree of correlation in the motion between the different protein domains and that the linker almost completely loses its correlation with the C-terminal domain, containing the active site tyrosine. CONCLUSIONS: These results indicate that a fully functional linker is required to confer camptothecin sensitivity to topoisomerase I since the destabilization of its structural-dynamical properties is correlated to an increase of religation rate and drug resistance.
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spelling pubmed-37667032013-09-09 Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico Tesauro, Cinzia Morozzo della Rocca, Blasco Ottaviani, Alessio Coletta, Andrea Zuccaro, Laura Arnò, Barbara D'Annessa, Ilda Fiorani, Paola Desideri, Alessandro Mol Cancer Research BACKGROUND: DNA topoisomerases are key enzymes that modulate the topological state of DNA through the breaking and rejoining of DNA strands. Human topoisomerase IB can be inhibited by several compounds that act through different mechanisms, including clinically used drugs, such as the derivatives of the natural compound camptothecin that reversibly bind the covalent topoisomerase-DNA complex, slowing down the religation of the cleaved DNA strand, thus inducing cell death. Three enzyme mutations, which confer resistance to irinotecan in an adenocarcinoma cell line, were recently identified but the molecular mechanism of resistance was unclear. METHODS: The three resistant mutants have been investigated in S. cerevisiae model system following their viability in presence of increasing amounts of camptothecin. A systematical analysis of the different catalytic steps has been made for one of these mutants (Glu710Gly) and has been correlated with its structural-dynamical properties studied by classical molecular dynamics simulation. RESULTS: The three mutants display a different degree of camptothecin resistance in a yeast cell viability assay. Characterization of the different steps of the catalytic cycle of the Glu710Gly mutant indicated that its resistance is related to a high religation rate that is hardly affected by the presence of the drug. Analysis of the dynamic properties through simulation indicate that the mutant displays a much lower degree of correlation in the motion between the different protein domains and that the linker almost completely loses its correlation with the C-terminal domain, containing the active site tyrosine. CONCLUSIONS: These results indicate that a fully functional linker is required to confer camptothecin sensitivity to topoisomerase I since the destabilization of its structural-dynamical properties is correlated to an increase of religation rate and drug resistance. BioMed Central 2013-09-03 /pmc/articles/PMC3766703/ /pubmed/24004603 http://dx.doi.org/10.1186/1476-4598-12-100 Text en Copyright © 2013 tesauro et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Tesauro, Cinzia
Morozzo della Rocca, Blasco
Ottaviani, Alessio
Coletta, Andrea
Zuccaro, Laura
Arnò, Barbara
D'Annessa, Ilda
Fiorani, Paola
Desideri, Alessandro
Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico
title Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico
title_full Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico
title_fullStr Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico
title_full_unstemmed Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico
title_short Molecular mechanism of the camptothecin resistance of Glu710Gly topoisomerase IB mutant analyzed in vitro and in silico
title_sort molecular mechanism of the camptothecin resistance of glu710gly topoisomerase ib mutant analyzed in vitro and in silico
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3766703/
https://www.ncbi.nlm.nih.gov/pubmed/24004603
http://dx.doi.org/10.1186/1476-4598-12-100
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