Identification of the consensus DNA sequence for Nczf binding.
The Nczf gene, which is identified as a target gene of Ncx, encodes a novel Kruppel-associated box (KRAB) zinc finger protein, which functions as a sequence-specific transcriptional repressor. We generated a fusion protein of the zinc finger domain of Nczf and glutathione S-transferase to identify Nczf-binding consensus DNA sequences with random oligonucleotides of 15 and 35 bases. The consensus binding sequence of core nucleotides contains (A/T/C)CTTT(A/G)TTNT. In a gel mobility shift assay, the probe containing these sequences bound to the fusion protein. In silico analysis, these consensus sequences were found on regulatory regions of the endothelin receptor B and the microphthalmia-associated transcription factor genes, which are involved in neural crest development. These results suggest that Nczf functions as a sequence-specific transcription repressor to regulate neural crest cell development.
Kitahashi
Masayasu
M
Department of Developmental Genetics, Graduate School of Medicine, Chiba University, Chiba, Japan.
Sato
Yoshiharu
Y
Fujimura
Lisa
L
Ozeki
Chisa
C
Arima
Masafumi
M
Sakamoto
Akemi
A
Yamamoto
Shuichi
S
Tokuhisa
Takeshi
T
Hatano
Masahiko
M
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism*; D016384:Consensus Sequence; D004247:DNA / Q000235:genetics* / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*; D012689:Sequence Homology, Nucleic Acid; D016335:Zinc Fingers / Q000235:genetics
D002352:Carrier Proteins; D018398:Homeodomain Proteins; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C107836:Tlx2 protein, mouse; D004247:DNA; C486788:Zfp472 protein, mouse