Sequence-specific DNA binding activity in the RAE28 protein, a mouse homologue of the Drosophila polyhomeotic protein.
The rae28 gene, a mouse homologue of the Drosophila polyhomeotic gene, is involved in the maintenance of the transcriptional repression states of Hox genes. In this study we synthesized the glutathione S transferase-RAE28 (GST-RAE28) fusion protein and examined sequence-specific DNA binding activity in the RAE28 protein by using the selected and amplified binding site method. After five rounds of enrichment, the eluted DNAs were amplified, cloned and sequenced. The sequences of individual oligonucleotides included the following consensus sequences; 5'-ACCA-3', 5'-ACCCA-3', 5'-CTATCA-3' and 5'-TGCC-3'. The oligonucleotides including these consensus sequences were show to have significant affinity with the GST-RAE28 fusion protein. The RAE28 protein was recently shown to form multimeric protein complexes with other members of mouse Pc-G proteins in the nucleus. These findings strongly suggest that the RAE28 protein constitutes a sequence-specific DNA binding domain in multimeric Pc-G protein complexes.
Nomura
M
M
Department of Medical Genetics, Research Institute for Microbial Diseases, Osaka University, Japan. shimada@biken.osaka-u.ac.jp
Takihara
Y
Y
Abdul Motaleb
M
M
Horie
K
K
Higashinakagawa
T
T
Shimada
K
K
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins; D016384:Consensus Sequence; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D015345:Oligonucleotide Probes / Q000378:metabolism; D063150:Polycomb Repressive Complex 1; D016133:Polymerase Chain Reaction; D011189:Potassium Chloride; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA
D002352:Carrier Proteins; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D015345:Oligonucleotide Probes; C103827:Phc1 protein, mouse; D011993:Recombinant Fusion Proteins; D011189:Potassium Chloride; D004247:DNA; D063150:Polycomb Repressive Complex 1