Affinity selection of DNA-binding protein complexes using mRNA display

Seiji Tateyama, Kenichi Horisawa, Hideaki Takashima, Etsuko Miyamoto-Sato, Nobuhide Doi, Hiroshi Yanagawa

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Comprehensive analysis of DNA-protein interactions is important for mapping transcriptional regulatory networks on a genome-wide level. Here we present a new application of mRNA display for in vitro selection of DNA-binding protein heterodimeric complexes. Under improved selection conditions using a TPA-responsive element (TRE) as a bait DNA, known interactors c-fos and c-jun were simultaneously enriched about 100-fold from a model library (a 1:1:20 000 mixture of c-fos, c-jun and gst genes) after one round of selection. Furthermore, almost all kinds of the AP-1 family genes including c-jun, c-fos, junD, junB, atf2 and b-atf were successfully selected from an mRNA display library constructed from a mouse brain poly A+ RNA after six rounds of selection. These results indicate that the mRNA display selection system can identify a variety of DNA-binding protein complexes in a single experiment. Since almost all transcription factors form heterooligomeric complexes to bind with their target DNA, this method should be most useful to search for DNA-binding transcription factor complexes.

Original languageEnglish
Article numbere27
JournalNucleic Acids Research
Volume34
Issue number3
DOIs
Publication statusPublished - 2006

Fingerprint

DNA-Binding Proteins
jun Genes
Messenger RNA
DNA
Transcription Factors
Gene Regulatory Networks
Transcription Factor AP-1
Libraries
Genome
Brain
Proteins

ASJC Scopus subject areas

  • Genetics

Cite this

Tateyama, S., Horisawa, K., Takashima, H., Miyamoto-Sato, E., Doi, N., & Yanagawa, H. (2006). Affinity selection of DNA-binding protein complexes using mRNA display. Nucleic Acids Research, 34(3), [e27]. https://doi.org/10.1093/nar/gnj025

Affinity selection of DNA-binding protein complexes using mRNA display. / Tateyama, Seiji; Horisawa, Kenichi; Takashima, Hideaki; Miyamoto-Sato, Etsuko; Doi, Nobuhide; Yanagawa, Hiroshi.

In: Nucleic Acids Research, Vol. 34, No. 3, e27, 2006.

Research output: Contribution to journalArticle

Tateyama, S, Horisawa, K, Takashima, H, Miyamoto-Sato, E, Doi, N & Yanagawa, H 2006, 'Affinity selection of DNA-binding protein complexes using mRNA display', Nucleic Acids Research, vol. 34, no. 3, e27. https://doi.org/10.1093/nar/gnj025
Tateyama, Seiji ; Horisawa, Kenichi ; Takashima, Hideaki ; Miyamoto-Sato, Etsuko ; Doi, Nobuhide ; Yanagawa, Hiroshi. / Affinity selection of DNA-binding protein complexes using mRNA display. In: Nucleic Acids Research. 2006 ; Vol. 34, No. 3.
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