title: Loss of responsiveness to chemotactic factors by deletion of the C-terminal protein interaction site of angiomotin.
abstract: We have recently identified a novel protein, named angiomotin, by its ability to bind the angiogenesis inhibitor angiostatin in the yeast two-hybrid system. Angiomotin belongs to a family with two other members, AmotL-1 and -2 characterized by coiled-coil and C-terminal PDZ binding domains. Here we show that the putative PDZ binding motif of angiomotin serves as a protein recognition site and that deletion of three amino acids in this site results in inhibition of chemotaxis. Furthermore, endothelial cells expressing mutant angiomotin failed to migrate and form tubes in an in vitro tube formation assay. To study the effect of angiomotin on embryonic angiogenesis, we generated transgenic mice expressing wild-type angiomotin and the C-terminal deletion mutant driven by the endothelial cell-specific receptor tyrosine kinase (TIE) promoter. Expression of mutant angiomotin in endothelial cells inhibited migration into the neuroectoderm and intersomitic regions resulting in death at embryonic day 9.5. In contrast, mice expressing wild-type angiomotin developed normally and were fertile. These results suggest that the putative PDZ binding motif of angiomotin plays a critical role in regulating the responsiveness of endothelial cells to chemotactic cues.
authors.affiliation: Department of Oncology-Pathology, Cancer Center Karolinska Institutet, S-17176 Stockholm, Sweden.
authors.forename: Tetyana
authors.initials: T
authors.lastname: Levchenko
authors.forename: Karin
authors.initials: K
authors.lastname: Aase
authors.forename: Boris
authors.initials: B
authors.lastname: Troyanovsky
authors.forename: Anders
authors.initials: A
authors.lastname: Bratt
authors.forename: Lars
authors.initials: L
authors.lastname: Holmgren
mesh_terms: D000091082:Angiomotins; D000091304:Angiopoietin-Like Protein 1; D043168:Angiostatins / Q000494:pharmacology; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D002633:Chemotaxis / Q000502:physiology*; D004622:Embryo, Mammalian / Q000166:cytology; D042783:Endothelial Cells / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D008840:Microfilament Proteins; D008856:Microscopy, Fluorescence; D009154:Mutation; D018919:Neovascularization, Physiologic / Q000187:drug effects; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D020798:Two-Hybrid System Techniques
chemical_list: C423985:AMOT protein, human; C510063:AMOTL1 protein, human; C449807:Amotl1 protein, mouse; D000091082:Angiomotins; D000091304:Angiopoietin-Like Protein 1; D002352:Carrier Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D008840:Microfilament Proteins; D043168:Angiostatins