English   español  
Por favor, use este identificador para citar o enlazar a este item: http://hdl.handle.net/10261/122677
Título

Disease-causing mutations in BEST1 gene are associated with altered sorting of bestrophin-1 protein

AutorDoumanov, Jordan A.; Domínguez Giménez, Paloma; Krishna, Abhay ; Bellido, María Luz ; Bhattacharya, Shom Shanker
Fecha de publicación22-jul-2013
EditorMultidisciplinary Digital Publishing Institute
CitaciónInternational Journal of Molecular Sciences 14(7): 15121- 15140 (2013)
ResumenMutations in BEST1 gene, encoding the bestrophin-1 (Best1) protein are associated with macular dystrophies. Best1 is predominantly expressed in the retinal pigment epithelium (RPE), and is inserted in its basolateral membrane. We investigated the cellular localization in polarized MDCKII cells of disease-associated Best1 mutant proteins to study specific sorting motifs of Best1. Real-time PCR and western blots for endogenous expression of BEST1 in MDCK cells were performed. Best1 mutant constructs were generated using site-directed mutagenesis and transfected in MDCK cells. For protein sorting, confocal microscopy studies, biotinylation assays and statistical methods for quantification of mislocalization were used. Analysis of endogenous expression of BEST1 in MDCK cells revealed the presence of BEST1 transcript but no protein. Confocal microscopy and quantitative analyses indicate that transfected normal human Best1 displays a basolateral localization in MDCK cells, while cell sorting of several Best1 mutants (Y85H, Q96R, L100R, Y227N, Y227E) was altered. In contrast to constitutively active Y227E, constitutively inactive Y227F Best1 mutant localized basolaterally similar to the normal Best1 protein. Our data suggest that at least three basolateral sorting motifs might be implicated in proper Best1 basolateral localization. In addition, non-phosphorylated tyrosine 227 could play a role for basolateral delivery. © 2013 by the authors; licensee MDPI, Basel, Switzerland.
DescripciónDoumanov, Jordan A. et al.
Versión del editorhttp://dx.doi.org/10.3390/ijms140715121
URIhttp://hdl.handle.net/10261/122677
DOI10.3390/ijms140715121
Identificadoresdoi: 10.3390/ijms140715121
issn: 1422-0067
Aparece en las colecciones: (CABIMER) Artículos
Ficheros en este ítem:
Fichero Descripción Tamaño Formato  
Disease_Causing_Mutations_Doumanov.pdf4,02 MBAdobe PDFVista previa
Visualizar/Abrir
Mostrar el registro completo
 

Artículos relacionados:


NOTA: Los ítems de Digital.CSIC están protegidos por copyright, con todos los derechos reservados, a menos que se indique lo contrario.