Expression of the Longin domain of TI-VAMP impairs lysosomal secretion and epithelial cell migration.
Biol Cell
; 99(5): 261-71, 2007 May.
Article
em En
| MEDLINE
| ID: mdl-17288539
ABSTRACT
BACKGROUND INFORMATION TI-VAMP (tetanus neurotoxin-insensitive vesicle-associated membrane protein; also called VAMP7) belongs to the Longin subfamily of v-SNAREs (vesicular soluble N-ethylmaleimide-sensitive fusion protein-attachment protein receptors). The regulatory N-terminal extension, called the Longin domain, of TI-VAMP has been shown previously to have a dual biochemical function it inhibits the capacity of TI-VAMP to form SNARE complexes and it binds to the delta subunit of the AP-3 (adaptor protein 3) complex in early endosomes, thereby targeting TI-VAMP to late endosomes. RESULTS:
We have generated MDCK (Madin-Darby canine kidney) cell lines expressing the Longin domain of TI-VAMP coupled to GFP (green fluorescent protein) in a doxycycline-dependent manner. As expected, AP-3delta (AP-3 delta subunit) is not properly localized in Longin-expressing cells. We have shown that the expression of the Longin domain impairs lysosomal secretion, as determined by the release of a pre-internalized fluorescent fluid-phase marker and by electron microscopy of the membrane-associated released particles. Membrane repair following mechanical wounding, a process requiring lysosomal secretion, is also impaired in cells expressing the Longin domain. Furthermore, cell migration, assessed by wound healing of MDCK monolayers, is also inhibited.CONCLUSIONS:
The results of the present study suggest that the expression of the Longin domain of TI-VAMP regulates lysosomal secretion of epithelial cells and provide molecular evidence for a role of the late endocytic system in cell migration.
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Base de dados:
MEDLINE
Assunto principal:
Expressão Gênica
/
Movimento Celular
/
Células Epiteliais
/
Proteínas R-SNARE
/
Lisossomos
Limite:
Animals
Idioma:
En
Revista:
Biol Cell
Ano de publicação:
2007
Tipo de documento:
Article
País de afiliação:
França