Cargando…

Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation

Protein kinase D (PKD) plays a critical role at the trans-Golgi network by regulating the fission of transport carriers destined for the plasma membrane. Two known Golgi-localized PKD substrates, PI4-kinase IIIβ and the ceramide transfer protein CERT, mediate PKD signaling to influence vesicle traff...

Descripción completa

Detalles Bibliográficos
Autores principales: Nhek, Sokha, Ngo, Mike, Yang, Xuemei, Ng, Michelle M., Field, Seth J., Asara, John M., Ridgway, Neale D., Toker, Alex
Formato: Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2893995/
https://www.ncbi.nlm.nih.gov/pubmed/20444975
http://dx.doi.org/10.1091/mbc.E10-02-0090
_version_ 1782183112823799808
author Nhek, Sokha
Ngo, Mike
Yang, Xuemei
Ng, Michelle M.
Field, Seth J.
Asara, John M.
Ridgway, Neale D.
Toker, Alex
author_facet Nhek, Sokha
Ngo, Mike
Yang, Xuemei
Ng, Michelle M.
Field, Seth J.
Asara, John M.
Ridgway, Neale D.
Toker, Alex
author_sort Nhek, Sokha
collection PubMed
description Protein kinase D (PKD) plays a critical role at the trans-Golgi network by regulating the fission of transport carriers destined for the plasma membrane. Two known Golgi-localized PKD substrates, PI4-kinase IIIβ and the ceramide transfer protein CERT, mediate PKD signaling to influence vesicle trafficking to the plasma membrane and sphingomyelin synthesis, respectively. PKD is recruited and activated at the Golgi through interaction with diacylglycerol, a pool of which is generated as a by-product of sphingomyelin synthesis from ceramide. Here we identify a novel substrate of PKD at the Golgi, the oxysterol-binding protein OSBP. Using a substrate-directed phospho-specific antibody that recognizes the optimal PKD consensus motif, we show that PKD phosphorylates OSBP at Ser240 in vitro and in cells. We further show that OSBP phosphorylation occurs at the Golgi. Phosphorylation of OSBP by PKD does not modulate dimerization, sterol binding, or affinity for PI(4)P. Instead, phosphorylation attenuates OSBP Golgi localization in response to 25-hydroxycholesterol and cholesterol depletion, impairs CERT Golgi localization, and promotes Golgi fragmentation.
format Text
id pubmed-2893995
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher The American Society for Cell Biology
record_format MEDLINE/PubMed
spelling pubmed-28939952010-09-16 Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation Nhek, Sokha Ngo, Mike Yang, Xuemei Ng, Michelle M. Field, Seth J. Asara, John M. Ridgway, Neale D. Toker, Alex Mol Biol Cell Articles Protein kinase D (PKD) plays a critical role at the trans-Golgi network by regulating the fission of transport carriers destined for the plasma membrane. Two known Golgi-localized PKD substrates, PI4-kinase IIIβ and the ceramide transfer protein CERT, mediate PKD signaling to influence vesicle trafficking to the plasma membrane and sphingomyelin synthesis, respectively. PKD is recruited and activated at the Golgi through interaction with diacylglycerol, a pool of which is generated as a by-product of sphingomyelin synthesis from ceramide. Here we identify a novel substrate of PKD at the Golgi, the oxysterol-binding protein OSBP. Using a substrate-directed phospho-specific antibody that recognizes the optimal PKD consensus motif, we show that PKD phosphorylates OSBP at Ser240 in vitro and in cells. We further show that OSBP phosphorylation occurs at the Golgi. Phosphorylation of OSBP by PKD does not modulate dimerization, sterol binding, or affinity for PI(4)P. Instead, phosphorylation attenuates OSBP Golgi localization in response to 25-hydroxycholesterol and cholesterol depletion, impairs CERT Golgi localization, and promotes Golgi fragmentation. The American Society for Cell Biology 2010-07-01 /pmc/articles/PMC2893995/ /pubmed/20444975 http://dx.doi.org/10.1091/mbc.E10-02-0090 Text en © 2010 by The American Society for Cell Biology
spellingShingle Articles
Nhek, Sokha
Ngo, Mike
Yang, Xuemei
Ng, Michelle M.
Field, Seth J.
Asara, John M.
Ridgway, Neale D.
Toker, Alex
Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation
title Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation
title_full Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation
title_fullStr Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation
title_full_unstemmed Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation
title_short Regulation of Oxysterol-binding Protein Golgi Localization through Protein Kinase D–mediated Phosphorylation
title_sort regulation of oxysterol-binding protein golgi localization through protein kinase d–mediated phosphorylation
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2893995/
https://www.ncbi.nlm.nih.gov/pubmed/20444975
http://dx.doi.org/10.1091/mbc.E10-02-0090
work_keys_str_mv AT nheksokha regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation
AT ngomike regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation
AT yangxuemei regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation
AT ngmichellem regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation
AT fieldsethj regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation
AT asarajohnm regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation
AT ridgwaynealed regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation
AT tokeralex regulationofoxysterolbindingproteingolgilocalizationthroughproteinkinasedmediatedphosphorylation