Spatial regulation of Drosophila snake protease activity in the generation of dorsal-ventral polarity.
Positional information along the dorsal-ventral axis of the Drosophila embryo is acquired through a signal transduction pathway which employs a extracellular protease cascade. The sequential activation of serine protease zymogens results in the ventrally localized production of a ligand in the perivitelline space of the embryo. Snake is one of several serine proteases which function in generating the ventralizing signal. Here, we investigate the biochemical properties of Snake in vivo and in vitro using recombinant forms of the protease. Wild-type Snake zymogen completely rescues embryos from snake null females when microinjected into the perivitelline space. Biochemical evidence for a covalently associated two-chain form of the activated protease is presented. The contribution of the activation peptide region to zymogen activation was addressed using site-directed mutagenesis. The phenotypic rescue properties of an autoactivated form of Snake reveal that the covalently associated proenzyme polypeptide chain suppresses a dominant effect associated with the activated catalytic chain alone. Recombinant active catalytic chain was produced and found to be short lived as a recombinant protein. These results suggest a model in which the proenzyme polypeptide both stabilizes and targets the Snake catalytic chain to a ventrally localized activation complex within the perivitelline space.
Smith
C L
CL
Molecular Biology Department, Sloan-Kettering Institute for Cancer Research, New York, NY 10021, USA.
Giordano
H
H
Schwartz
M
M
DeLotto
R
R
D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D004330:Drosophila / Q000196:embryology* / Q000235:genetics; D029721:Drosophila Proteins; D004789:Enzyme Activation; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction
D000911:Antibodies, Monoclonal; D029721:Drosophila Proteins; D015345:Oligonucleotide Probes; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C081508:snk protein, Drosophila