DEV00558 Data Supplement

Files in this Data Supplement:

• Figure S1 -

  Fig. S1. (A) Similarities between the Boz-eh1 motif (amino acids 9 to 15) and those of various other repressor proteins. Amino acids that deviate from Boz are highlighted in blue. D.r., Danio rerio; X.l., Xenopus laevis; D.m., Drosophila melanogaster; M.m., Mus musculus. Gsc (Ferreiro et al., 1998), Hex (Ho et al., 1999), Xvent1 (Friedle et al., 1998; Rastegar et al., 1999), Vox (Melby et al., 1999), Xanf1 (Ermakova et al., 1999) and Xvent2 (Trindade et al., 1999). (B) A reporter assay to determine whether Boz acts as a transcriptional activator or repressor in yeast. The integrated GAL1-lacZ reporter gene contains five upstream binding sites for GAL4 and one for GCN4. As yeast cells lack endogenous GAL4, this reporter gene is activated by endogenous GCN4. b-Galactosidase activity was determined in yeast cells containing plasmids that express either no GAL4 protein (pMA200), the GAL4 DNA-binding domain [GAL4(1-147)(pMA241)], a fusion of the GAL4 DNA binding domain with the full-length Boz protein [GAL4(1-147)-Boz], or two well-characterized yeast transcriptional repressors [GAL4(1-147)-SSB24 and GAL4(1-147)-SSB16].

  REFERENCES

  Ermakova, G. V., Alexandrova, E. M., Kazanskaya, O. V., Vasiliev, O. L., Smith, M. W. and Zaraisky, A. G. (1999). The homeobox gene, Xanf-1, can control both neural differentiation and patterning in the presumptive anterior neurectoderm of the Xenopus laevis embryo. Development 126, 4513-4523.

  Ferreiro, B., Artinger, M., Cho, K. and Niehrs, C. (1998). Antimorphic goosecoids. Development 125, 1347-1359.

  Friedle, H., Rastegar, S., Paul, H., Kaufmann, E. and Knöchel, W. (1998). Xvent-1 mediates BMP-4-induced suppression of the dorsal-lip-specific early response gene XFD-1' in Xenopus embryos. EMBO J. 17, 2298-2307.

  Ho, C. Y., Houart, C., Wilson, S. W. and Stainier, D. Y. (1999). A role for the extraembryonic yolk syncytial layer in patterning the zebrafish embryo suggested by properties of the hex gene. Curr. Biol. 9, 1131-1134.

  Melby, A. E., Clements, W. K. and Kimelman, D. (1999). Regulation of dorsal gene expression in Xenopus by the ventralizing homeodomain gene Vox. Dev. Biol. 211, 293-305.

  Rastegar, S., Friedle, H., Frommer, G. and Knöchel, W. (1999). Transcriptional regulation of Xvent homeobox genes. Mech. Dev. 81, 139-149.

  Trindade, M., Tada, M. and Smith, J. C. (1999). DNA-binding specificity and embryological function of Xom (Xvent-2). Dev. Biol. 216, 442-456.

• Figure S2 - Fig. S2. A bcd fusion gene was generated that drives expression of Boz from a localized mRNA at the anterior tip of the Drosophila egg. (A-D) Cuticle phenotype of (A) wild-type larvae, (B) larvae from bcd^E1 homozygous mutant females, (C) an embryo from a wild-type female carrying one copy of the bcd-boz transgene and (D) an embryo from a bcd^E1/TM3 female carrying one copy of the bcd-boz transgene. Regardless of whether the females were wild type or heterozygous mutant for bcd, embryos from females with a single bcd-boz transgene express a dominant phenotype that resembles that of bcd. (E,F) Expression of hunchback (hb) during syncytial blastoderm stages in embryos from wild-type females (E) and from females carrying one copy of the bcd-boz fusion gene (F). (G,H) Expression of even-skipped (eve) during syncytial blastoderm stages in embryos from wild-type females (G) and from females carrying one copy of the bcd-boz fusion gene (H). Fluorescent (Cy3) anti-Eve immunohistochemistry reveals segmentation defects similar to those observed in embryos from bcd mutant females. A-H are lateral views, anterior towards the left and dorsal towards the top.