Ca2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination.

Handle URI:
http://hdl.handle.net/10754/596774
Title:
Ca2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination.
Authors:
Fornander, Louise H; Frykholm, Karolin; Reymer, Anna; Renodon-Cornière, Axelle; Takahashi, Masayuki; Nordén, Bengt
Abstract:
Human RAD51 protein (HsRad51) catalyses the DNA strand exchange reaction for homologous recombination. To clarify the molecular mechanism of the reaction in vitro being more effective in the presence of Ca(2+) than of Mg(2+), we have investigated the effect of these ions on the structure of HsRad51 filament complexes with single- and double-stranded DNA, the reaction intermediates. Flow linear dichroism spectroscopy shows that the two ionic conditions induce significantly different structures in the HsRad51/single-stranded DNA complex, while the HsRad51/double-stranded DNA complex does not demonstrate this ionic dependence. In the HsRad51/single-stranded DNA filament, the primary intermediate of the strand exchange reaction, ATP/Ca(2+) induces an ordered conformation of DNA, with preferentially perpendicular orientation of nucleobases relative to the filament axis, while the presence of ATP/Mg(2+), ADP/Mg(2+) or ADP/Ca(2+) does not. A high strand exchange activity is observed for the filament formed with ATP/Ca(2+), whereas the other filaments exhibit lower activity. Molecular modelling suggests that the structural variation is caused by the divalent cation interfering with the L2 loop close to the DNA-binding site. It is proposed that the larger Ca(2+) stabilizes the loop conformation and thereby the protein-DNA interaction. A tight binding of DNA, with bases perpendicularly oriented, could facilitate strand exchange.
Citation:
Fornander LH, Frykholm K, Reymer A, Renodon-Corniere A, Takahashi M, et al. (2012) Ca2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination. Nucleic Acids Research 40: 4904–4913. Available: http://dx.doi.org/10.1093/nar/gks140.
Publisher:
Oxford University Press (OUP)
Journal:
Nucleic Acids Research
KAUST Grant Number:
KUK-11-008-23
Issue Date:
22-Feb-2012
DOI:
10.1093/nar/gks140
PubMed ID:
22362735
PubMed Central ID:
PMC3367181
Type:
Article
ISSN:
0305-1048; 1362-4962
Sponsors:
King Abdullah University of Science and Technology Grant (KUK-11-008-23 to L.H.F., K.F. and A.R.); European Research Council (ERC-2008-AdG 227700 to B.N.); Agence Nationale de la Recherche Grant (ANR-2010-BLAN-1013 DynRec to A.R.-C. and M.T.); BiogenOuest to A.R.-C. and M.T. Funding for open access charge: Chalmers Library.
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Full metadata record

DC FieldValue Language
dc.contributor.authorFornander, Louise Hen
dc.contributor.authorFrykholm, Karolinen
dc.contributor.authorReymer, Annaen
dc.contributor.authorRenodon-Cornière, Axelleen
dc.contributor.authorTakahashi, Masayukien
dc.contributor.authorNordén, Bengten
dc.date.accessioned2016-02-21T08:50:24Zen
dc.date.available2016-02-21T08:50:24Zen
dc.date.issued2012-02-22en
dc.identifier.citationFornander LH, Frykholm K, Reymer A, Renodon-Corniere A, Takahashi M, et al. (2012) Ca2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination. Nucleic Acids Research 40: 4904–4913. Available: http://dx.doi.org/10.1093/nar/gks140.en
dc.identifier.issn0305-1048en
dc.identifier.issn1362-4962en
dc.identifier.pmid22362735en
dc.identifier.doi10.1093/nar/gks140en
dc.identifier.urihttp://hdl.handle.net/10754/596774en
dc.description.abstractHuman RAD51 protein (HsRad51) catalyses the DNA strand exchange reaction for homologous recombination. To clarify the molecular mechanism of the reaction in vitro being more effective in the presence of Ca(2+) than of Mg(2+), we have investigated the effect of these ions on the structure of HsRad51 filament complexes with single- and double-stranded DNA, the reaction intermediates. Flow linear dichroism spectroscopy shows that the two ionic conditions induce significantly different structures in the HsRad51/single-stranded DNA complex, while the HsRad51/double-stranded DNA complex does not demonstrate this ionic dependence. In the HsRad51/single-stranded DNA filament, the primary intermediate of the strand exchange reaction, ATP/Ca(2+) induces an ordered conformation of DNA, with preferentially perpendicular orientation of nucleobases relative to the filament axis, while the presence of ATP/Mg(2+), ADP/Mg(2+) or ADP/Ca(2+) does not. A high strand exchange activity is observed for the filament formed with ATP/Ca(2+), whereas the other filaments exhibit lower activity. Molecular modelling suggests that the structural variation is caused by the divalent cation interfering with the L2 loop close to the DNA-binding site. It is proposed that the larger Ca(2+) stabilizes the loop conformation and thereby the protein-DNA interaction. A tight binding of DNA, with bases perpendicularly oriented, could facilitate strand exchange.en
dc.description.sponsorshipKing Abdullah University of Science and Technology Grant (KUK-11-008-23 to L.H.F., K.F. and A.R.); European Research Council (ERC-2008-AdG 227700 to B.N.); Agence Nationale de la Recherche Grant (ANR-2010-BLAN-1013 DynRec to A.R.-C. and M.T.); BiogenOuest to A.R.-C. and M.T. Funding for open access charge: Chalmers Library.en
dc.publisherOxford University Press (OUP)en
dc.rightsThis is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.en
dc.rights.urihttp://creativecommons.org/licenses/by-nc/3.0en
dc.titleCa2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination.en
dc.typeArticleen
dc.identifier.journalNucleic Acids Researchen
dc.identifier.pmcidPMC3367181en
dc.contributor.institutionChalmers University of Technology, Göteborg, Swedenen
dc.contributor.institutionGoteborgs Universitet, Goteborg, Swedenen
dc.contributor.institutionUniversite de Nantes, Nantes, Franceen
kaust.grant.numberKUK-11-008-23en

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