TY - JOUR
T1 - Affinity selection of DNA-binding protein complexes using mRNA display
AU - Tateyama, Seiji
AU - Horisawa, Kenichi
AU - Takashima, Hideaki
AU - Miyamoto-Sato, Etsuko
AU - Doi, Nobuhide
AU - Yanagawa, Hiroshi
N1 - Funding Information:
The authors thank Ms A. Utsumi for experimental support. This work was supported in part by a Grant-in-Aid for Scientific Research, and a Special Coordination Fund from the Ministry of Education, Culture, Sports, Science and Technology, Japan.
PY - 2006
Y1 - 2006
N2 - 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.
AB - 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.
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U2 - 10.1093/nar/gnj025
DO - 10.1093/nar/gnj025
M3 - Article
C2 - 16478713
AN - SCOPUS:33644863777
SN - 0305-1048
VL - 34
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 3
M1 - e27
ER -