Biological activity of soluble wingless protein in cultured Drosophila imaginal disc cells

THE phenotypes caused by mutations in Wnt genes suggest that their gene products are involved in cell-to-cell communication1–6. Wnt genes indeed encode secreted molecules7–10, but soluble active Wnt protein has not been found. We have developed a novel cell culture assay for the Drosophila Wnt gene wingless 11,12, using a Drosophila imaginal disc cell line (cl-8; ref. 13), and measured effects on the adherens junction protein armadillo14,15, a known genetic target of wingless 16. Transfection of a temperature-sensitive wingless complementary DNA into cl-8 cells increases the levels of the armadillo protein. The wingless protein does not affect the rate of synthesis of armadillo, but leads to increased stability of an otherwise rapidly decaying armadillo protein. The wingless protein in the extracellular matrix and soluble medium from donor cells also increases the levels of armadillo protein. The protein in the medium acts fast and is inhibited by an antibody to wingless protein, demonstrating that Wnt products can act as soluble extracellular signalling molecules.

[1]  M. Peifer,et al.  The segment polarity gene armadillo encodes a functionally modular protein that is the Drosophila homolog of human plakoglobin , 1990, Cell.

[2]  W. Talbot,et al.  The drosophila EcR gene encodes an ecdysone receptor, a new member of the steroid receptor superfamily , 1991, Cell.

[3]  E. Wieschaus,et al.  wingless signal and Zeste-white 3 kinase trigger opposing changes in the intracellular distribution of Armadillo. , 1994, Development.

[4]  Detlef Weigel,et al.  The Drosophila homology of the mouse mammary oncogene int-1 is identical to the segment polarity gene wingless , 1987, Cell.

[5]  A. Martinez Arias,et al.  Roles of wingless in patterning the larval epidermis of Drosophila. , 1991, Development.

[6]  M. Krasnow,et al.  Intercellular signalling in Drosophila segment formation reconstructed in vitro , 1993, Nature.

[7]  P. Lawrence,et al.  Distribution of the wingless gene product in drosophila embryos: A protein involved in cell-cell communication , 1989, Cell.

[8]  A. M. Arias,et al.  Secretion and movement of wingless protein in the epidermis of the Drosophila embryo , 1991, Mechanisms of Development.

[9]  N. Perrimon,et al.  Mutations in the segment polarity genes wingless and porcupine impair secretion of the wingless protein. , 1993, The EMBO journal.

[10]  C W Turck,et al.  A homolog of the armadillo protein in Drosophila (plakoglobin) associated with E-cadherin. , 1991, Science.

[11]  Norbert Perrimon,et al.  Putative protein kinase product of the Drosophila segment-polarity gene zeste-white3 , 1990, Nature.

[12]  A. McMahon The Wnt family of developmental regulators , 1992 .

[13]  M. Peifer,et al.  The product of the Drosophila segment polarity gene armadillo is part of a multi-protein complex resembling the vertebrate adherens junction. , 1993, Journal of cell science.

[14]  Norbert Perrimon,et al.  Components of wingless signalling in Drosophila , 1994, Nature.

[15]  C. Doe,et al.  Neuroblast specification and formation regulated by wingless in the Drosophila CNS. , 1993, Science.

[16]  Konrad Basler,et al.  Organizing activity of wingless protein in Drosophila , 1993, Cell.

[17]  B. Schryver,et al.  Secreted int-1 protein is associated with the cell surface. , 1990, Molecular and cellular biology.

[18]  N E Baker,et al.  Molecular cloning of sequences from wingless, a segment polarity gene in Drosophila: the spatial distribution of a transcript in embryos , 1987, The EMBO journal.

[19]  A. Joyner,et al.  The midbrain-hindbrain phenotype of Wnt-1− Wnt-1− mice results from stepwise deletion of engrailed-expressing cells by 9.5 days postcoitum , 1992, Cell.

[20]  P. Simpson,et al.  An early embryonic product of the gene shaggy encodes a serine/threonine protein kinase related to the CDC28/cdc2+ subfamily. , 1990, The EMBO journal.

[21]  A. Brown,et al.  The proto‐oncogene int‐1 encodes a secreted protein associated with the extracellular matrix. , 1990, The EMBO journal.

[22]  Judith A. Kassis,et al.  Two-tiered regulation of spatially patterned engrailed gene expression during Drosophila embryogenesis , 1988, Nature.

[23]  E. Wieschaus,et al.  Spatial expression of the Drosophila segment polarity gene armadillo is posttranscriptionally regulated by wingless , 1990, Cell.

[24]  Norbert Perrimon,et al.  dishevelled and armadillo act in the Wingless signalling pathway in Drosophila , 1994, Nature.