RESUMEN
Angiostatin is a potent inhibitor of angiogenesis. One mechanism through which angiostatin inhibits angiogenesis is by binding to the cell surface protein p80-angiomotin. The p80-angiomotin protein promotes angiogenesis, in part, by conferring a hypermigratory phenotype to endothelial cells. Although p80-angiomotin is extensively characterized, less is known about the related protein angiomotin-like 1. We report that angiomotin-like 1 forms part of a protein complex containing p80-angiomotin. Structure-function studies revealed that angiomotin-like 1 associates with this p80-angiomotin-containing complex via its coiled-coil domain. Since p80-angiomotin plays a role in cell migration, a process that involves the remodeling of the actin cytoskeleton, we then addressed the hypothesis that angiomotin-like 1 may interact with the cytoskeleton. Immunofluorescence studies reveal that angiomotin-like 1 not only co-localizes with filamentous actin but also significantly modifies the architecture of the actin cytoskeleton. Regarding migration, angiomotin-like 1 increases the velocity of migration and decreases the persistence of migration directionality. Together these observations strongly suggest that angiomotin-like 1 is involved in actin-cytoskeleton-based processes, in part, via its interaction with a p80-angiomotin-containing complex and the actin cytoskeleton. These findings have important implications for angiogenesis-driven disease since angiomotin and angiomotin-like 1 are both expressed in capillaries.
Asunto(s)
Actinas/metabolismo , Citoesqueleto/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Proteínas de la Membrana/metabolismo , Neovascularización Fisiológica , Angiomotinas , Movimiento Celular/fisiología , Células HeLa , Humanos , Proteínas de la Membrana/genética , Proteínas de Microfilamentos , Estructura Terciaria de Proteína/genética , Estructura Terciaria de Proteína/fisiologíaRESUMEN
The motin family of proteins is comprised of three polypeptides, angiomotin, angiomotin-like 1, and angiomotin-like 2. Angiomotin is an angiostatin-binding protein that promotes endothelial cell motility and is involved in angiogenesis. The function of human angiomotin-like-1 and angiomotin-like-2, however, remains unknown. In this report, we investigated the potential for molecular diversity within the motin family of proteins through the identification and characterization of alternatively spliced transcripts in endothelial cells, human tissues and a variety of cell lines. We report that the motins display variability at the mRNA level suggesting an intricate regulatory system at the transcriptional and potentially protein level. Some alternative transcripts are expressed in a tissue-specific manner and others give rise to novel protein isoforms. The alternative splicing that occurs within this protein family may have important implications in the regulation of the expression and function of the motins.