Zebrafish Pax9 encodes two proteins with distinct C-terminal transactivating domains of different potency negatively regulated by adjacent N-terminal sequences.
We describe the isolation of cDNA clones for zebrafish Pax9. Pax9 expression was initiated at the end of the segmentation period in mesenchymal sclerotome cells on both sides of the notochord similarly to the corresponding mouse and chick genes. Two transcripts, Pax9a and -b, are generated by alternative splicing. The gene contains 4 exons with exon 3 being included in the Pax9a transcript and spliced out in the Pax9b transcript. The Pax9a and -b proteins are identical for 212 amino acids from the N terminus but contain distinct C-terminal regions of 131 and 58 amino acids, respectively. The paired domain of Pax9 displayed a binding-site specificity distinct from Pax6 but similar to Pax1 and -2. Both Pax9a and -b activated a promoter containing a paired domain binding site. However, this activation was observed when low amounts of Pax9 expression vectors were used. Higher amounts led to a sharp decrease in the activation and even turned into repression. Both the distinct C-terminal regions of Pax9a and -b harbored transcriptional activating domains of different potency not revealed in the context of the full-length proteins due to a negative influence of the N-terminal region including the paired domain.
Departments of Biochemistry, Institute of Medical Biology, University of Tromso, 9037 Tromso, Norway.
S
S
Nornes
I
I
Mikkola
S
S
Krauss
M
M
Delghandi
M
M
Perander
T
T
Johansen
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D019077:Helix-Turn-Helix Motifs; D006801:Humans; D008969:Molecular Sequence Data; D051764:PAX9 Transcription Factor; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*; D015533:Transcriptional Activation; D015027:Zebrafish / Q000196:embryology; D029961:Zebrafish Proteins
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D051764:PAX9 Transcription Factor; D014157:Transcription Factors; D029961:Zebrafish Proteins; C497324:pax9 protein, zebrafish