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Schnölzer, Martina
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article
The Wnt-specific astacin proteinase HAS-7 restricts head organizer formation in Hydra
Abstract
<jats:title>Abstract</jats:title><jats:sec><jats:title>Background</jats:title><jats:p>The <jats:italic>Hydra</jats:italic> head organizer acts as a signaling center that initiates and maintains the primary body axis in steady state polyps and during budding or regeneration. Wnt/beta-Catenin signaling functions as a primary cue controlling this process, but how Wnt ligand activity is locally restricted at the protein level is poorly understood. Here we report a proteomic analysis of <jats:italic>Hydra</jats:italic> head tissue leading to the identification of an astacin family proteinase as a Wnt processing factor.</jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p><jats:italic>Hydra</jats:italic> astacin-7 (HAS-7) is expressed from gland cells as an apical-distal gradient in the body column, peaking close beneath the tentacle zone. <jats:italic>HAS-7</jats:italic> siRNA knockdown abrogates HyWnt3 proteolysis in the head tissue and induces a robust double axis phenotype, which is rescued by simultaneous <jats:italic>HyWnt3</jats:italic> knockdown. Accordingly, double axes are also observed in conditions of increased Wnt activity as in transgenic actin::HyWnt3 and <jats:italic>HyDkk1/2/4</jats:italic> siRNA treated animals. HyWnt3-induced double axes in <jats:italic>Xenopus</jats:italic> embryos could be rescued by coinjection of <jats:italic>HAS-7</jats:italic> mRNA. Mathematical modelling combined with experimental promotor analysis indicate an indirect regulation of <jats:italic>HAS-7</jats:italic> by beta-Catenin, expanding the classical Turing-type activator-inhibitor model.</jats:p></jats:sec><jats:sec><jats:title>Conclusions</jats:title><jats:p>We show the astacin family protease HAS-7 maintains a single head organizer through proteolysis of HyWnt3. Our data suggest a negative regulatory function of Wnt processing astacin proteinases in the global patterning of the oral-aboral axis in <jats:italic>Hydra</jats:italic>.</jats:p></jats:sec>