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Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level

Human topoisomerase I (hTopI) is an essential cellular enzyme. The enzyme is often upregulated in cancer cells, and it is a target for chemotherapeutic drugs of the camptothecin (CPT) family. Response to CPT-based treatment is dependent on hTopI activity, and reduction in activity, and mutations in...

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Autores principales: Proszek, Joanna, Roy, Amit, Jakobsen, Ann-Katrine, Frøhlich, Rikke, Knudsen, Birgitta R., Stougaard, Magnus
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3926610/
https://www.ncbi.nlm.nih.gov/pubmed/24434877
http://dx.doi.org/10.3390/s140101195
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author Proszek, Joanna
Roy, Amit
Jakobsen, Ann-Katrine
Frøhlich, Rikke
Knudsen, Birgitta R.
Stougaard, Magnus
author_facet Proszek, Joanna
Roy, Amit
Jakobsen, Ann-Katrine
Frøhlich, Rikke
Knudsen, Birgitta R.
Stougaard, Magnus
author_sort Proszek, Joanna
collection PubMed
description Human topoisomerase I (hTopI) is an essential cellular enzyme. The enzyme is often upregulated in cancer cells, and it is a target for chemotherapeutic drugs of the camptothecin (CPT) family. Response to CPT-based treatment is dependent on hTopI activity, and reduction in activity, and mutations in hTopI have been reported to result in CPT resistance. Therefore, hTOPI gene copy number, mRNA level, protein amount, and enzyme activity have been studied to explain differences in cellular response to CPT. We show that Rolling Circle Enhanced Enzyme Activity Detection (REEAD), allowing measurement of hTopI cleavage-religation activity at the single molecule level, may be used to detect posttranslational enzymatic differences influencing CPT response. These differences cannot be detected by analysis of hTopI gene copy number, mRNA amount, or protein amount, and only become apparent upon measuring the activity of hTopI in the presence of CPT. Furthermore, we detected differences in the activity of the repair enzyme tyrosyl-DNA phosphodiesterase 1, which is involved in repair of hTopI-induced DNA damage. Since increased TDP1 activity can reduce cellular CPT sensitivity we suggest that a combined measurement of TDP1 activity and hTopI activity in presence of CPT will be the best determinant for CPT response.
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spelling pubmed-39266102014-02-18 Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level Proszek, Joanna Roy, Amit Jakobsen, Ann-Katrine Frøhlich, Rikke Knudsen, Birgitta R. Stougaard, Magnus Sensors (Basel) Article Human topoisomerase I (hTopI) is an essential cellular enzyme. The enzyme is often upregulated in cancer cells, and it is a target for chemotherapeutic drugs of the camptothecin (CPT) family. Response to CPT-based treatment is dependent on hTopI activity, and reduction in activity, and mutations in hTopI have been reported to result in CPT resistance. Therefore, hTOPI gene copy number, mRNA level, protein amount, and enzyme activity have been studied to explain differences in cellular response to CPT. We show that Rolling Circle Enhanced Enzyme Activity Detection (REEAD), allowing measurement of hTopI cleavage-religation activity at the single molecule level, may be used to detect posttranslational enzymatic differences influencing CPT response. These differences cannot be detected by analysis of hTopI gene copy number, mRNA amount, or protein amount, and only become apparent upon measuring the activity of hTopI in the presence of CPT. Furthermore, we detected differences in the activity of the repair enzyme tyrosyl-DNA phosphodiesterase 1, which is involved in repair of hTopI-induced DNA damage. Since increased TDP1 activity can reduce cellular CPT sensitivity we suggest that a combined measurement of TDP1 activity and hTopI activity in presence of CPT will be the best determinant for CPT response. Molecular Diversity Preservation International (MDPI) 2014-01-10 /pmc/articles/PMC3926610/ /pubmed/24434877 http://dx.doi.org/10.3390/s140101195 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Proszek, Joanna
Roy, Amit
Jakobsen, Ann-Katrine
Frøhlich, Rikke
Knudsen, Birgitta R.
Stougaard, Magnus
Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level
title Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level
title_full Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level
title_fullStr Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level
title_full_unstemmed Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level
title_short Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level
title_sort topoisomerase i as a biomarker: detection of activity at the single molecule level
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3926610/
https://www.ncbi.nlm.nih.gov/pubmed/24434877
http://dx.doi.org/10.3390/s140101195
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