Expression, localization and functional role of small GTPases of the Rab3 family in insulin-secreting cells

Détails

ID Serval
serval:BIB_8EE599BB8E55
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Expression, localization and functional role of small GTPases of the Rab3 family in insulin-secreting cells
Périodique
Journal of Cell Science
Auteur(s)
Regazzi  R., Ravazzola  M., Iezzi  M., Lang  J., Zahraoui  A., Andereggen  E., Morel  P., Takai  Y., Wollheim  C. B.
ISSN
0021-9533 (Print)
Statut éditorial
Publié
Date de publication
09/1996
Volume
109 ( Pt 9)
Pages
2265-73
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Sep
Résumé
We examined the presence of small molecular mass GTP-binding proteins of the Rab3 family in different insulin-secreting cells. Rab3B and Rab3C were identified by western blotting in rat and in human pancreatic islets, in two rat insulin-secreting cell lines, RINm5F and INS-1, as well as in the hamster cell line HIT-T15. In contrast, Rab3A was detected in rat pancreatic islets as well as in the two insulin-secreting rat cell lines but not in human pancreatic islets and was only barely discernible in HIT-T15 cells. These findings were confirmed by two-dimensional gel electrophoresis followed by GTP-overlay of homogenates of pancreatic islets and of the purified protein. Northern blotting analysis revealed that Rab3D is expressed in the same insulin-secreting cells as Rab3A. Separation of the cells of the rat islets by fluorescence-activated cell sorting demonstrated that Rab3A was exclusively expressed in beta-cells. Rab3A was found to be associated with insulin-containing secretory granules both by immunofluorescence, immunoelectron microscopy and after sucrose density gradient. Overexpression in HIT-T15 cells of a Rab3A mutant deficient in GTP hydrolysis inhibited insulin secretion stimulated by a mixture of nutrients and bombesin. Insulin release triggered by these secretagogues was also slightly decreased by the overexpression of wild-type Rab3A but not by the overexpression of wild-type Rab5A and of a Rab5A mutant deficient in GTP hydrolysis. Finally, we studied the expression in insulin-secreting cells of rabphilin-3A, a putative effector protein that associates with the GTP-bound form of Rab3A. This Rab3A effector was not detectable in any of the cells investigated in the present study. Taken together these results indicate an involvement of Rab3A in the control of insulin release in rat and hamster. In human beta-cells, a different Rab3 isoform but with homologous function may replace Rab3A.
Mots-clé
Animals Cell Line Cricetinae GTP-Binding Proteins/*genetics/metabolism/physiology Gene Expression Guanosine Triphosphate/metabolism Humans Hydrolysis Immunohistochemistry Insulin/*secretion Islets of Langerhans/cytology/*metabolism/secretion Molecular Weight Mutation RNA, Messenger/genetics/metabolism Rats Species Specificity Subcellular Fractions/metabolism Transfection rab3 GTP-Binding Proteins
Pubmed
Web of science
Création de la notice
24/01/2008 15:30
Dernière modification de la notice
03/03/2018 19:19
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