Post-endocytotic Deubiquitination and Degradation of the MetabotropicAminobutyric Acid Receptor by the Ubiquitin-specific Protease 14

0471850 - ÚMG 2017 RIV US eng J - Článek v odborném periodiku
Lahaie, N. - Králíková, Michaela - Prezeau, L. - Blahoš, Jaroslav - Bouvier, M.
Post-endocytotic Deubiquitination and Degradation of the MetabotropicAminobutyric Acid Receptor by the Ubiquitin-specific Protease 14.
Journal of Biological Chemistry. Roč. 291, č. 13 (2016), s. 7156-7170. ISSN 0021-9258. E-ISSN 1083-351X
Grant CEP: GA ČR GAP303/12/2408
Institucionální podpora: RVO:68378050
Klíčová slova: bioluminescence resonance energy transfer (BRET) * biosensor * deubiquitylation * G protein-coupled receptor (GPCR) * GABA receptor * protein degradation * receptor endocytosis * ubiquitination
Kód oboru RIV: EB - Genetika a molekulární biologie
Impakt faktor: 4.125, rok: 2016

Mechanisms controlling the metabotropicaminobutyric acid receptor (GABA(B)) cell surface stability are still poorly understood. In contrast with many other G protein-coupled receptors (GPCR), it is not subject to agonist-promoted internalization, but is constitutively internalized and rapidly down-regulated. In search of novel interacting proteins regulating receptor fate, we report that the ubiquitin-specific protease 14 (USP14) interacts with the GABA(B(1b)) subunit's second intracellular loop. Probing the receptor for ubiquitination using bioluminescence resonance energy transfer (BRET), we detected a constitutive and phorbol 12-myristate 13-acetate (PMA)-induced ubiquitination of the receptor at the cell surface. PMA also increased internalization and accelerated receptor degradation. Overexpression of USP14 decreased ubiquitination while treatment with a small molecule inhibitor of the deubiquitinase (IU1) increased receptor ubiquitination. Treatment with the internalization inhibitor Dynasore blunted both USP14 and IU1 effects on the receptor ubiquitination state, suggesting a post-endocytic site of action. Overexpression of USP14 also led to an accelerated degradation of GABA(B) in a catalytically independent fashion. We thus propose a model whereby cell surface ubiquitination precedes endocytosis, after which USP14 acts as an ubiquitin-binding protein that targets the ubiquitinated receptor to lysosomal degradation and promotes its deubiquitination.
Trvalý link: http://hdl.handle.net/11104/0269208