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dc.contributor.authorKing, Gordon J
dc.contributor.authorChanson, Aurelie H
dc.contributor.authorMcCallum, Emily J
dc.contributor.authorOhme-Takagi, Masaru
dc.contributor.authorByriel, Karl
dc.contributor.authorHill, Justine M
dc.contributor.authorMartin, Jennifer L
dc.contributor.authorMylne, Joshua S
dc.date.accessioned2018-01-24T01:52:50Z
dc.date.available2018-01-24T01:52:50Z
dc.date.issued2013
dc.identifier.issn0021-9258
dc.identifier.doi10.1074/jbc.M112.438572
dc.identifier.urihttp://hdl.handle.net/10072/171962
dc.description.abstractThe B3 DNA-binding domain is a plant-specific domain found throughout the plant kingdom from the alga Chlamydomonas to grasses and flowering plants. Over 100 B3 domain-containing proteins are found in the model plant Arabidopsis thaliana, and one of these is critical for accelerating flowering in response to prolonged cold treatment, an epigenetic process called vernalization. Despite the specific phenotype of genetic vrn1 mutants, the VERNALIZATION1 (VRN1) protein localizes throughout the nucleus and shows sequence-nonspecific binding in vitro. In this work, we used a dominant repressor tag that overcomes genetic redundancy to show that VRN1 is involved in processes beyond vernalization that are essential for Arabidopsis development. To understand its sequence-nonspecific binding, we crystallized VRN1(208–341) and solved its crystal structure to 1.6 Å resolution using selenium/single-wavelength anomalous diffraction methods. The crystallized construct comprises the second VRN1 B3 domain and a preceding region conserved among VRN1 orthologs but absent in other B3 domains. We established the DNA-binding face using NMR and then mutated positively charged residues on this surface with a series of 16 Ala and Glu substitutions, ensuring that the protein fold was not disturbed using heteronuclear single quantum correlation NMR spectra. The triple mutant R249E/R289E/R296E was almost completely incapable of DNA binding in vitro. Thus, we have revealed that although VRN1 is sequence-nonspecific in DNA binding, it has a defined DNA-binding surface.
dc.description.peerreviewedYes
dc.languageEnglish
dc.language.isoeng
dc.publisherThe American Society for Biochemistry and Molecular Biology Inc
dc.relation.ispartofpagefrom3198
dc.relation.ispartofpageto3207
dc.relation.ispartofissue5
dc.relation.ispartofjournalJournal of Biological Chemistry
dc.relation.ispartofvolume288
dc.subject.fieldofresearchChemical sciences
dc.subject.fieldofresearchBiological sciences
dc.subject.fieldofresearchBiochemistry and cell biology not elsewhere classified
dc.subject.fieldofresearchBiomedical and clinical sciences
dc.subject.fieldofresearchcode34
dc.subject.fieldofresearchcode31
dc.subject.fieldofresearchcode310199
dc.subject.fieldofresearchcode32
dc.titleThe Arabidopsis B3 domain protein VERNALIZATION1 (VRN1) is involved in processes essential for development, with structural and mutational studies revealing its DNA-binding surface
dc.typeJournal article
dc.type.descriptionC1 - Articles
dc.type.codeC - Journal Articles
gro.hasfulltextNo Full Text
gro.griffith.authorMartin, Jennifer


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