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dc.contributor.authorStotz, Henrik
dc.contributor.authorFindling, Simone
dc.contributor.authorNukarinen, Ella
dc.contributor.authorWeckwerth, Wolfram
dc.contributor.authorMueller, Martin J.
dc.contributor.authorBerger, Susanne
dc.date.accessioned2017-03-30T15:00:56Z
dc.date.available2017-03-30T15:00:56Z
dc.date.issued2014-12-22
dc.identifier.citationStotz , H , Findling , S , Nukarinen , E , Weckwerth , W , Mueller , M J & Berger , S 2014 , ' A tandem affinity purification tag of TGA2 for isolation of interacting proteins in Arabidopsis thaliana ' , Plant Signaling and Behavior , vol. 9 , no. 10 , e972794 . https://doi.org/10.4161/15592316.2014.972794
dc.identifier.issn1559-2316
dc.identifier.otherPURE: 8442320
dc.identifier.otherPURE UUID: 7f5992d4-b7a9-4980-bc13-94f7dd476870
dc.identifier.otherPubMed: 25482810
dc.identifier.otherScopus: 84922318837
dc.identifier.urihttp://hdl.handle.net/2299/17739
dc.descriptionThis is an Accepted Manuscript of an article published by Taylor & Francis Group in Plant Signaling & Behavior, on 31 October 2014, available online: https://doi.org/10.4161/15592316.2014.972794.
dc.description.abstractTandem affinity purification (TAP) tagging provides a powerful tool for isolating interacting proteins in vivo. TAP-tag purification offers particular advantages for the identification of stimulus-induced protein interactions. Type II bZIP transcription factors (TGA2, TGA5 and TGA6) play key roles in pathways that control salicylic acid, ethylene, xenobiotic and reactive oxylipin signaling. Although proteins interacting with these transcription factors have been identified through genetic and yeast 2-hybrid screening, others are still elusive. We have therefore generated a C-terminal TAP-tag of TGA2 to isolate additional proteins that interact with this transcription factor. Three lines most highly expressing TAP-tagged TGA2 were functional in that they partially complemented reactive oxylipin-responsive gene expression in a tga2 tga5 tga6 triple mutant. TAP-tagged TGA2 in the most strongly overexpressing line was proteolytically less stable than in the other 2 lines. Only this overexpressing line could be used in a 2-step purification process, resulting in isolation of co-purifying bands of larger molecular weight than TGA2. TAP-tagged TGA2 was used to pull down NPR1, a protein known to interact with this transcription factor. Mass spectrometry was used to identify peptides that co-purified with TAP-tagged TGA2. Having generated this TGA2 TAP-tag line will therefore be an asset to researchers interested in stimulus-induced signal transduction processes.en
dc.language.isoeng
dc.relation.ispartofPlant Signaling and Behavior
dc.subject12-oxo-phytodienoic acid
dc.subjectlutathione-S-transferase
dc.subjectlipid stress
dc.subjectprotein complex
dc.subjectthale cress
dc.titleA tandem affinity purification tag of TGA2 for isolation of interacting proteins in Arabidopsis thalianaen
dc.contributor.institutionDepartment of Human and Environmental Sciences
dc.contributor.institutionSchool of Life and Medical Sciences
dc.contributor.institutionHealth & Human Sciences Research Institute
dc.contributor.institutionAgriculture, Food and Veterinary Sciences
dc.contributor.institutionGeography, Environment and Agriculture
dc.contributor.institutionCrop Protection and Climate Change
dc.description.statusPeer reviewed
rioxxterms.versionAM
rioxxterms.versionofrecordhttps://doi.org/10.4161/15592316.2014.972794
rioxxterms.typeJournal Article/Review
herts.preservation.rarelyaccessedtrue


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