ArticleBiochemistry2008
Purification and identification of positive regulators binding to a novel element in the c-Jun promoter.
Article in Biochemistry, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 19 citations in OpenAlex.
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- Phosphorylation of ATF2 and interaction with NFY induces c-Jun in the gonadotrope.Molecular and cellular endocrinology · 2013Article
- Unbiased proteomic analysis of proteins interacting with the HIV-1 5'LTR sequence: role of the transcription factor Meis.Nucleic acids research · 2012Article
- Asymmetric polymerase chain reaction provides alternatives for preparation of (GT)₅-tailed duplex DNA promoter for promoter trapping.Analytical biochemistry · 2012Article
- Transcription factor proteomics: identification by a novel gel mobility shift-three-dimensional electrophoresis method coupled with southwestern blot and high-performance liquid chromatography-electrospray-mass spectrometry analysis.Journal of chromatography. A · 2011Article
- CCR5 promoter haplotype transcription complex characterization.Journal of health care for the poor and underserved · 2011Article
- Repeated probing of Southwestern blots using alkaline phosphatase stripping.Journal of chromatography. A · 2010Article
- Article
- Methods for proteomic analysis of transcription factors.Journal of chromatography. A · 2009Review
- Two-dimensional southwestern blotting and characterization of transcription factors on-blot.Journal of proteome research · 2009Article
- Ku proteins interact with activator protein-2 transcription factors and contribute to ERBB2 overexpression in breast cancer cell lines.Breast cancer research : BCR · 2009Article
- Purification and characterization of transcription factors.Mass spectrometry reviewsReview
- Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
Abstract
A putative response element, GAGCCTC, was observed years ago in footprinting analysis of the c-jun promoter, and here we investigate its function in regulating c-jun expression and identify a protein complex that binds there. Electrophoretic mobility shift assays demonstrate a sequence-specific binding complex with this element in HEK293 cells. Additionally, unlabeled consensus AP-1 element DNA, but not a similar NF-jun element DNA, competes with complex formation. Mutations of this element decrease c-jun promoter reporter activity by nearly 5-fold in HEK293 cells. A new, two-step oligonucleotide trapping technique was developed to purify the element binding proteins. LC-nanospray-ESI-MS/MS identification and Western blotting show that the purified complex contains Ku80 and c-jun, which was further confirmed by antibody supershift, by immunoprecipitation with Southwestern blot or with UV cross-linking analysis in vitro as well as chromatin immunoprecipitation in vivo. c-Jun promoter activity and c-jun expression were decreased by Ku80 siRNA introduction. A mutant Ku80 plasmid with normal amino acid sequence but immune to the siRNA recovers c-jun promoter activity from siRNA inhibition. Similarly, Ku70 wild type transfection can also upregulate c-jun promoter activity. Thus, Ku80-c-jun activates c-jun expression by binding to this GAGCCTC element in the c-jun promoter and Ku70 may also serve a role.
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