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Vol. 14, No. 10, pp. 1279-1289, May 15, 2000
Departments of 1 Biochemistry and Biophysics, and
Physiology Programs in Developmental Biology, Genetics
and Human Genetics, University of California, San Francisco (UCSF), San
Francisco, California 94143-0448 USA
Vertebrate endoderm development has recently become the focus of
intense investigation. In this report, we first show that the zebrafish
bonnie and clyde (bon) gene plays a critical early role
in endoderm formation. bon mutants exhibit a profound reduction in the number of sox17-expressing endodermal precursors formed during gastrulation, and, consequently, a profound reduction in gut
tissue at later stages. The endodermal precursors that do form in
bon mutants, however, appear to differentiate normally indicating that bon is not required at later steps of endoderm development. We further demonstrate that bon encodes a
paired-class homeodomain protein of the Mix family that is expressed
transiently before and during early gastrulation in both mesodermal and
endodermal progenitors. Overexpression of bon can rescue
endodermal gene expression and the formation of a gut tube in
bon mutants. Analysis of a newly identified mutant allele
reveals that a single amino acid substitution in the DNA recognition
helix of the homeodomain creates a dominant interfering form of Bon
when overexpressed. We also show through loss- and gain-of-function
analyses that Bon functions exclusively downstream of cyclops
and squint signaling. Together, our data demonstrate that Bon
is a critical transcriptional regulator of early endoderm formation.
[Key Words: Paired-class homeodomain; sox17; cyclops; squint; gut]
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