Biochemical characterization of the Drosophila dpp protein, a member of the transforming growth factor beta family of growth factors.
The decapentaplegic (dpp) gene of Drosophila melanogaster is required for pattern formation in the embryo and for viability of the epithelial cells in the imaginal disks. The dpp protein product predicted from the DNA sequence is similar to members of a family of growth factors that includes transforming growth factor beta (TGF-beta). We have produced polyclonal antibodies to a recombinant dpp protein made in bacteria and used a metallothionein promoter to express a dpp cDNA in Drosophila S2 cells. Similar to other proteins in the TGF-beta family, the dpp protein produced by the Drosophila cells was proteolytically cleaved, and both portions of the protein were secreted from the cells. The amino-terminal 47-kilodalton (kDa) peptide was found in the medium and in the proteins adhering to the plastic petri dish. The carboxy-terminal peptide, the region with sequence similarity to the active ligand portion of TGF-beta, was found extracellularly as a 30-kDa homodimer. Most of the 30-kDa homodimer was in the S2 cell protein adsorbed onto the surface of the plastic dish. The dpp protein could be released into solution by increased salt concentration and nonionic detergent. Under these conditions, the amino-terminal and carboxy-terminal portions of dpp were not associated in a stable complex.
Panganiban
G E
GE
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Rashka
K E
KE
Neitzel
M D
MD
Hoffmann
F M
FM
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002999:Clone Cells; D029721:Drosophila Proteins; D004331:Drosophila melanogaster / Q000235:genetics*; D006133:Growth Substances / Q000235:genetics; D007301:Insect Hormones / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids; D012313:RNA / Q000302:isolation & purification; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D015291:Transforming Growth Factors / Q000235:genetics*
D029721:Drosophila Proteins; D006133:Growth Substances; D007301:Insect Hormones; D046911:Macromolecular Substances; C064196:dpp protein, Drosophila; D012313:RNA; D015291:Transforming Growth Factors