Identification of tissue-specific DNA-protein binding sites by means of two-dimensional electrophoretic mobility shift assay display.
We developed a technique of differential electrophoretic mobility shift assay (EMSA) display allowing identification of tissue-specific protein-binding sites within long genomic sequences. Using this approach, we identified 10 cell type-specific protein-binding sites (protein target sites [PTSs]) within a 137-kb human chromosome 19 region. In general, tissue-specific binding of proteins from different nuclear extracts by individual PTSs did not follow the all-or-nothing principle. Most often, PTS-protein complexes were formed in all cases, but they were different for different nuclear extracts used.
Chernov
Igor P
IP
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia.
Timchenko
Kira A
KA
Akopov
Sergey B
SB
Nikolaev
Lev G
LG
Sverdlov
Eugene D
ED
D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002874:Chromosome Mapping / Q000379:methods*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D003626:Data Display; D004587:Electrophoresis, Agar Gel; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay / Q000295:instrumentation / Q000379:methods*; D015723:Gene Library; D005821:Genetic Techniques; D006801:Humans; D007525:Isoelectric Focusing; D009687:Nuclear Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D014018:Tissue Distribution; D014157:Transcription Factors / Q000737:chemistry*
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors