Bix1, a direct target of Xenopus T-box genes, causes formation of ventral mesoderm and endoderm.
Brachyury, a member of the T-box gene family, is required for posterior mesoderm and notochord differentiation in vertebrate development, and mis-expression of Xenopus Brachyury causes ectopic mesoderm formation. Brachyury is a transcription activator, and its ability to activate transcription is essential for its biological function, but Brachyury target genes have proved difficult to identify. Here we employ a hormone-inducible Brachyury construct and subtractive hybridization to search for such targets. Using this approach we have isolated Bix1, a homeobox gene expressed both in the marginal zone of Xenopus and in the vegetal hemisphere. Expression of Bix1 is induced in an immediate-early fashion by mesoderm-inducing factors such as activin as well as by the products of the T-box genes Xbra and VegT (also known as Antipodean, Brat and Xombi). Activation of Bix1 in response to Xbra is direct in the sense that it does not require protein synthesis, and both Xbra and VegT activate expression of a reporter gene driven by the Bix 5' regulatory region, which contains an Xbra/VegT binding site. Mis-expression of low levels of Bix1 causes formation of ventral mesoderm, while high levels induce endodermal differentiation. These results suggest that Bix1 acts downstream of both VegT and Xbra to induce formation of mesoderm and endoderm.
Tada
M
M
Division of Developmental Biology, National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
Casey
E S
ES
Fairclough
L
L
Smith
J C
JC
D028341:Activins; D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002454:Cell Differentiation; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004627:Embryonic Induction; D004707:Endoderm / Q000378:metabolism*; D005326:Fetal Proteins; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D005801:Genes, Homeobox; D017781:Genes, Immediate-Early / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D007150:Immunohistochemistry; D007265:Inhibins / Q000235:genetics; D008648:Mesoderm / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D020218:Response Elements / Q000235:genetics; D020825:T-Box Domain Proteins; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D029867:Xenopus Proteins; D014982:Xenopus laevis / Q000196:embryology / Q000235:genetics; D000098908:Brachyury Protein
D028341:Activins; D004268:DNA-Binding Proteins; D005326:Fetal Proteins; D018398:Homeodomain Proteins; D007265:Inhibins; D012333:RNA, Messenger; D020825:T-Box Domain Proteins; D014157:Transcription Factors; D029867:Xenopus Proteins; D000098908:Brachyury Protein; C115868:Bix1 protein, Xenopus; C103966:VegT protein, Xenopus