Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures

Topological variants of single-strand DNA (ssDNA) structures, referred to as “functional DNA,” have been detected in regulatory regions of many genes and are thought to affect gene expression. Two fluorescent analog...

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Main Authors: Yogini P. Bhavsar, Samantha M. Reilly, Randy M. Wadkins
Format: Article
Language:English
Published: Wiley 2011-01-01
Series:Journal of Nucleic Acids
Online Access:http://dx.doi.org/10.4061/2011/986820
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author Yogini P. Bhavsar
Samantha M. Reilly
Randy M. Wadkins
author_facet Yogini P. Bhavsar
Samantha M. Reilly
Randy M. Wadkins
author_sort Yogini P. Bhavsar
collection DOAJ
description Topological variants of single-strand DNA (ssDNA) structures, referred to as “functional DNA,” have been detected in regulatory regions of many genes and are thought to affect gene expression. Two fluorescent analogs of deoxycytidine, Pyrrolo-dC (PdC) and 1,3-diaza-2-oxophenoxazine (tC∘), can be incorporated into DNA. Here, we describe spectroscopic studies of both analogs to determine fluorescent properties that report on structural transitions from double-strand DNA (dsDNA) to ssDNA, a common pathway in the transition to functional DNA structures. We obtained fluorescence-detected circular dichroism (FDCD) spectra, steady-state fluorescence spectra, and fluorescence lifetimes of the fluorophores in DNA. Our results show that PdC is advantageous in fluorescence lifetime studies because of a distinct ~2 ns change between paired and unpaired bases. However, tC∘ is a better probe for FDCD experiments that report on the helical structure of DNA surrounding the fluorophore. Both fluorophores provide complementary data to measure DNA structural transitions.
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spelling doaj-art-1aca5d73bc8e469595facfbc5d011dbd2025-02-03T01:27:35ZengWileyJournal of Nucleic Acids2090-021X2011-01-01201110.4061/2011/986820986820Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional StructuresYogini P. Bhavsar0Samantha M. Reilly1Randy M. Wadkins2Department of Chemistry and Biochemistry, The University of Mississippi, Coulter Hall, Room 409, MS 38677, USADepartment of Chemistry and Biochemistry, The University of Mississippi, Coulter Hall, Room 409, MS 38677, USADepartment of Chemistry and Biochemistry, The University of Mississippi, Coulter Hall, Room 409, MS 38677, USATopological variants of single-strand DNA (ssDNA) structures, referred to as “functional DNA,” have been detected in regulatory regions of many genes and are thought to affect gene expression. Two fluorescent analogs of deoxycytidine, Pyrrolo-dC (PdC) and 1,3-diaza-2-oxophenoxazine (tC∘), can be incorporated into DNA. Here, we describe spectroscopic studies of both analogs to determine fluorescent properties that report on structural transitions from double-strand DNA (dsDNA) to ssDNA, a common pathway in the transition to functional DNA structures. We obtained fluorescence-detected circular dichroism (FDCD) spectra, steady-state fluorescence spectra, and fluorescence lifetimes of the fluorophores in DNA. Our results show that PdC is advantageous in fluorescence lifetime studies because of a distinct ~2 ns change between paired and unpaired bases. However, tC∘ is a better probe for FDCD experiments that report on the helical structure of DNA surrounding the fluorophore. Both fluorophores provide complementary data to measure DNA structural transitions.http://dx.doi.org/10.4061/2011/986820
spellingShingle Yogini P. Bhavsar
Samantha M. Reilly
Randy M. Wadkins
Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures
Journal of Nucleic Acids
title Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures
title_full Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures
title_fullStr Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures
title_full_unstemmed Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures
title_short Evaluation of Fluorescent Analogs of Deoxycytidine for Monitoring DNA Transitions from Duplex to Functional Structures
title_sort evaluation of fluorescent analogs of deoxycytidine for monitoring dna transitions from duplex to functional structures
url http://dx.doi.org/10.4061/2011/986820
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AT randymwadkins evaluationoffluorescentanalogsofdeoxycytidineformonitoringdnatransitionsfromduplextofunctionalstructures