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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...
Autores principales: | , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2010
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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 |
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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 |
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