Xtbx6r, a novel T-box gene expressed in the paraxial mesoderm, has anterior neural-inducing activity.
T-box proteins are important transcriptional regulators in animal development. We searched the Xenopus laevis expressed sequence tag (EST) database using zebrafish tbx24 (Ztbx24) as a query and found a sequence. We then obtained corresponding clones from a neurula cDNA library. This novel gene has a T-box showing 53% homology with Xenopus laevis tbx6 (Xtbx6) and 51% with Ztbx24 at the amino acid level and is relatively close to Xtbx6 indicated by alignment analysis. In situ hybridization showed that it is expressed in the paraxial mesoderm in the caudal region in a very similar manner to Xtbx6, with a slight difference in that the former is emphasized more dorsally and has a more restricted distribution along the antero-posterior axis. From these results we named this gene Xtbx6r (tbx6-related). Xtbx6r or Xtbx6r-EnR, when overexpressed in animal caps, induced anterior neural markers but not mesodermal markers. In contrast, Xtbx6r-VP16, Xtbx6 and Ztbx24 induced various mesodermal markers. These results indicate that Xtbx6r is a transcriptional repressor and has activity different from that of Xtbx6 or Ztbx24. Xtbx6r induced Otx2, XAG and Pax6 in animal caps. This activity differed from that of Xbra-EnR or Xtbx6-EnR, suggesting that some differences in biological activity exist among the tested repressor-type T-box genes. Depletion of Xtbx6r by antisense morpholino oligo produced curved embryos, but did not affect expressions of MyoD, Myf5, XWnt8 or thylacine2, nor inhibited muscle differentiation or segmentation. The results of knockdown and overexpression experiments suggest that Xtbx6r is involved in some morphogenesis in the paraxial mesoderm.
Yabe
Shigeharu
S
Graduate School of Integrated Science, Yokohama City University, Kanazawa-Ku, Yokohama, Japan.
Tazumi
Shunsuke
S
Yokoyama
Jun
J
Uchiyama
Hideho
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004475:Ectoderm / Q000378:metabolism; D020224:Expressed Sequence Tags; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D008648:Mesoderm / Q000378:metabolism*; D008969:Molecular Sequence Data; D009474:Neurons / Q000378:metabolism*; D010957:Plasmids / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D020825:T-Box Domain Proteins / Q000096:biosynthesis / Q000235:genetics*; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics*; D029867:Xenopus Proteins / Q000096:biosynthesis / Q000235:genetics*; D014982:Xenopus laevis / Q000378:metabolism*
D020825:T-Box Domain Proteins; C491181:Tbx6 protein, Xenopus; C517392:Tbx6r protein, Xenopus; D014157:Transcription Factors; D029867:Xenopus Proteins