A novel paired domain DNA recognition motif can mediate Pax2 repression of gene transcription.
The paired domain (PD) is an evolutionarily conserved DNA-binding domain encoded by the Pax gene family of developmental regulators. The Pax proteins are transcription factors and are involved in a variety of processes such as brain development, patterning of the central nervous system (CNS), and B-cell development. In this report we demonstrate that the zebrafish Pax2 PD can interact with a novel type of DNA sequences in vitro, the triple-A motif, consisting of a heptameric nucleotide sequence G/CAAACA/TC with an invariant core of three adjacent adenosines. This recognition sequence was found to be conserved in known natural Pax5 repressor elements involved in controlling the expression of the p53 and J-chain genes. By identifying similar high affinity binding sites in potential target genes of the Pax2 protein, including the pax2 gene itself, we obtained further evidence that the triple-A sites are biologically significant. The putative natural target sites also provide a basis for defining an extended consensus recognition sequence. In addition, we observed in transformation assays a direct correlation between Pax2 repressor activity and the presence of triple-A sites. The results suggest that a transcriptional regulatory function of Pax proteins can be modulated by PD binding to different categories of target sequences.
Håvik
B
B
Department of Molecular Biology, University of Bergen, Bergen, N-5020, Norway.
Ragnhildstveit
E
E
Lorens
J B
JB
Saelemyr
K
K
Fauske
O
O
Knudsen
L K
LK
Fjose
A
A
D020121:5' Untranslated Regions / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002460:Cell Line; D003001:Cloning, Molecular; D016384:Consensus Sequence; D017124:Conserved Sequence; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D051762:PAX2 Transcription Factor; D010446:Peptide Fragments / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D015027:Zebrafish; D029961:Zebrafish Proteins
D020121:5' Untranslated Regions; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D051762:PAX2 Transcription Factor; D010446:Peptide Fragments; D012333:RNA, Messenger; D012097:Repressor Proteins; D014157:Transcription Factors; D029961:Zebrafish Proteins; C497320:pax2a protein, zebrafish