title: Application of mRNA display for in vitro selection of DNA-binding transcription factor complexes.
abstract: Comprehensive analysis of DNA-protein interactions is important for mapping transcriptional regulatory networks at the genome-wide level. Here, we present a new application of mRNA display, using the in vitro virus (IVV) technology, for in vitro selection of DNA-binding protein complexes. Under optimal selection conditions using bait DNAs, many kinds of DNA-binding protein complexes can be successfully selected from an mRNA display library constructed from poly A(+) RNA after several rounds of selection. This mRNA display selection system can identify a variety of DNA-binding protein complexes in a single experiment. Remarkably, this system can also select DNA-binding protein heterooligomeric complexes. Since almost all transcription factors form heterooligomeric complexes to bind with their target DNA, this method should be broadly useful to identify DNA-binding transcription factor complexes.
authors: Department of Biosciences and Informatics, Keio University, Yokohama, Japan.
authors: Seiji
authors: S
authors: Tateyama
authors: Hiroshi
authors: H
authors: Yanagawa
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002474:Cell-Free System; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D055767:Immobilized Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D025941:Protein Interaction Mapping / Q000379:methods*; D011691:Puromycin / Q000737:chemistry; D012333:RNA, Messenger / Q000737:chemistry*; D060888:Real-Time Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D003432:Cross-Linking Reagents; D017931:DNA Primers; D055767:Immobilized Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D011691:Puromycin; D004247:DNA