Facile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.

We have studied the structure adopted by an (A-T)34 sequence from a Xenopus globin gene when present in a negatively supercoiled plasmid. A variety of enzyme and chemical probing experiments and electrophoretic migration shift methods reveal that the sequence adopts cruciform geometry at moderate le...

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Main Authors: Greaves, D, Patient, R, Lilley, D
Format: Journal article
Language:English
Published: 1985
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author Greaves, D
Patient, R
Lilley, D
author_facet Greaves, D
Patient, R
Lilley, D
author_sort Greaves, D
collection OXFORD
description We have studied the structure adopted by an (A-T)34 sequence from a Xenopus globin gene when present in a negatively supercoiled plasmid. A variety of enzyme and chemical probing experiments and electrophoretic migration shift methods reveal that the sequence adopts cruciform geometry at moderate levels of supercoiling. The structure has the lowest free energy of formation yet observed for a cruciform, and no detectable kinetic barrier preventing rapid interconversion between extruded and unextruded conformations. Analysis of band-shift experiments reveals a twist change on cruciform formation of -5.8, slightly smaller than the -6.5 we would predict on the basis of a transition from B DNA. An attractive explanation consistent with this discrepancy is that the (A-T)34 stretch is locally underwound to about 11.7 base-pairs/helical turn at low levels of supercoiling. This calculation is made on the assumption that the cruciform junction is structurally similar to those examined previously, which is supported by the nuclease digestion results. This perturbed helical structure could be of considerable biological significance.
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spelling oxford-uuid:f0cd09cf-9945-4d89-a765-d2e7d129477d2022-03-27T11:50:59ZFacile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:f0cd09cf-9945-4d89-a765-d2e7d129477dEnglishSymplectic Elements at Oxford1985Greaves, DPatient, RLilley, DWe have studied the structure adopted by an (A-T)34 sequence from a Xenopus globin gene when present in a negatively supercoiled plasmid. A variety of enzyme and chemical probing experiments and electrophoretic migration shift methods reveal that the sequence adopts cruciform geometry at moderate levels of supercoiling. The structure has the lowest free energy of formation yet observed for a cruciform, and no detectable kinetic barrier preventing rapid interconversion between extruded and unextruded conformations. Analysis of band-shift experiments reveals a twist change on cruciform formation of -5.8, slightly smaller than the -6.5 we would predict on the basis of a transition from B DNA. An attractive explanation consistent with this discrepancy is that the (A-T)34 stretch is locally underwound to about 11.7 base-pairs/helical turn at low levels of supercoiling. This calculation is made on the assumption that the cruciform junction is structurally similar to those examined previously, which is supported by the nuclease digestion results. This perturbed helical structure could be of considerable biological significance.
spellingShingle Greaves, D
Patient, R
Lilley, D
Facile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.
title Facile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.
title_full Facile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.
title_fullStr Facile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.
title_full_unstemmed Facile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.
title_short Facile cruciform formation by an (A-T)34 sequence from a Xenopus globin gene.
title_sort facile cruciform formation by an a t 34 sequence from a xenopus globin gene
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