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Improved nuclear localization of DNA-binding polyamides
Regulation of endogenous genes by DNA-binding polyamides requires effective nuclear localization. Previous work employing confocal microscopy to study uptake of fluorophore-labeled polyamides has demonstrated the difficulty of predicting a priori the nuclear uptake of a given polyamide. The data sug...
Autores principales: | , , , |
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Formato: | Texto |
Lenguaje: | English |
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Oxford University Press
2007
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1802595/ https://www.ncbi.nlm.nih.gov/pubmed/17175539 http://dx.doi.org/10.1093/nar/gkl1042 |
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author | Nickols, Nicholas G. Jacobs, Claire S. Farkas, Michelle E. Dervan, Peter B. |
author_facet | Nickols, Nicholas G. Jacobs, Claire S. Farkas, Michelle E. Dervan, Peter B. |
author_sort | Nickols, Nicholas G. |
collection | PubMed |
description | Regulation of endogenous genes by DNA-binding polyamides requires effective nuclear localization. Previous work employing confocal microscopy to study uptake of fluorophore-labeled polyamides has demonstrated the difficulty of predicting a priori the nuclear uptake of a given polyamide. The data suggest that dye identity influences uptake sufficiently such that a dye-conjugate cannot be used as a proxy for unlabeled analogs. Polyamides capable of nuclear localization unaided by fluorescent dyes are desirable due to size and other limitations of fluorophores. Recently, a polyamide-fluorescein conjugate targeted to the hypoxia response element (HRE) was found to inhibit vascular endothelial growth factor (VEGF) expression in cultured HeLa cells. The current study uses inhibition of VEGF expression as a biological read-out for effective nuclear localization of HRE-targeted polyamides. We synthesized a focused library of non-fluorescent, HRE-targeted polyamides in which the C-terminus ‘tail’ has been systematically varied. Members of this library bind the HRE with affinities comparable or superior to that of the fluorescein-labeled analog. Although most library members demonstrate modest or no biological activity, two non-fluorescent polyamides are reported with activity rivaling that of the previously reported fluorescein-labeled polyamide. We also show evidence that promoter occupancy by HIF-1, the transcription factor that binds the HRE, is inhibited by HRE-targeted polyamides. |
format | Text |
id | pubmed-1802595 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2007 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-18025952007-03-01 Improved nuclear localization of DNA-binding polyamides Nickols, Nicholas G. Jacobs, Claire S. Farkas, Michelle E. Dervan, Peter B. Nucleic Acids Res Molecular Biology Regulation of endogenous genes by DNA-binding polyamides requires effective nuclear localization. Previous work employing confocal microscopy to study uptake of fluorophore-labeled polyamides has demonstrated the difficulty of predicting a priori the nuclear uptake of a given polyamide. The data suggest that dye identity influences uptake sufficiently such that a dye-conjugate cannot be used as a proxy for unlabeled analogs. Polyamides capable of nuclear localization unaided by fluorescent dyes are desirable due to size and other limitations of fluorophores. Recently, a polyamide-fluorescein conjugate targeted to the hypoxia response element (HRE) was found to inhibit vascular endothelial growth factor (VEGF) expression in cultured HeLa cells. The current study uses inhibition of VEGF expression as a biological read-out for effective nuclear localization of HRE-targeted polyamides. We synthesized a focused library of non-fluorescent, HRE-targeted polyamides in which the C-terminus ‘tail’ has been systematically varied. Members of this library bind the HRE with affinities comparable or superior to that of the fluorescein-labeled analog. Although most library members demonstrate modest or no biological activity, two non-fluorescent polyamides are reported with activity rivaling that of the previously reported fluorescein-labeled polyamide. We also show evidence that promoter occupancy by HIF-1, the transcription factor that binds the HRE, is inhibited by HRE-targeted polyamides. Oxford University Press 2007-01 2006-12-14 /pmc/articles/PMC1802595/ /pubmed/17175539 http://dx.doi.org/10.1093/nar/gkl1042 Text en © 2006 The Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Molecular Biology Nickols, Nicholas G. Jacobs, Claire S. Farkas, Michelle E. Dervan, Peter B. Improved nuclear localization of DNA-binding polyamides |
title | Improved nuclear localization of DNA-binding polyamides |
title_full | Improved nuclear localization of DNA-binding polyamides |
title_fullStr | Improved nuclear localization of DNA-binding polyamides |
title_full_unstemmed | Improved nuclear localization of DNA-binding polyamides |
title_short | Improved nuclear localization of DNA-binding polyamides |
title_sort | improved nuclear localization of dna-binding polyamides |
topic | Molecular Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1802595/ https://www.ncbi.nlm.nih.gov/pubmed/17175539 http://dx.doi.org/10.1093/nar/gkl1042 |
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