2'-Alkynyl spin-labelling is a minimally perturbing tool for DNA structural analysis.

Abstract:

:The determination of distances between specific points in nucleic acids is essential to understanding their behaviour at the molecular level. The ability to measure distances of 2-10 nm is particularly important: deformations arising from protein binding commonly fall within this range, but the reliable measurement of such distances for a conformational ensemble remains a significant challenge. Using several techniques, we show that electron paramagnetic resonance (EPR) spectroscopy of oligonucleotides spin-labelled with triazole-appended nitroxides at the 2' position offers a robust and minimally perturbing tool for obtaining such measurements. For two nitroxides, we present results from EPR spectroscopy, X-ray crystal structures of B-form spin-labelled DNA duplexes, molecular dynamics simulations and nuclear magnetic resonance spectroscopy. These four methods are mutually supportive, and pinpoint the locations of the spin labels on the duplexes. In doing so, this work establishes 2'-alkynyl nitroxide spin-labelling as a minimally perturbing method for probing DNA conformation.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Hardwick JS,Haugland MM,El-Sagheer AH,Ptchelkine D,Beierlein FR,Lane AN,Brown T,Lovett JE,Anderson EA

doi

10.1093/nar/gkaa086

subject

Has Abstract

pub_date

2020-04-06 00:00:00

pages

2830-2840

issue

6

eissn

0305-1048

issn

1362-4962

pii

5735211

journal_volume

48

pub_type

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