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Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments
The role of specific membrane lipids in transport between endoplasmic reticulum (ER) and Golgi compartments is poorly understood. Using cell-free assays that measure stages in ER-to-Golgi transport, we screened a variety of enzyme inhibitors, lipid-modifying enzymes, and lipid ligands to investigate...
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Formato: | Texto |
Lenguaje: | English |
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The American Society for Cell Biology
2011
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3020917/ https://www.ncbi.nlm.nih.gov/pubmed/21119004 http://dx.doi.org/10.1091/mbc.E10-04-0317 |
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author | Lorente-Rodríguez, Andrés Barlowe, Charles |
author_facet | Lorente-Rodríguez, Andrés Barlowe, Charles |
author_sort | Lorente-Rodríguez, Andrés |
collection | PubMed |
description | The role of specific membrane lipids in transport between endoplasmic reticulum (ER) and Golgi compartments is poorly understood. Using cell-free assays that measure stages in ER-to-Golgi transport, we screened a variety of enzyme inhibitors, lipid-modifying enzymes, and lipid ligands to investigate requirements in yeast. The pleckstrin homology (PH) domain of human Fapp1, which binds phosphatidylinositol-4-phosphate (PI(4)P) specifically, was a strong and specific inhibitor of anterograde transport. Analysis of wild type and mutant PH domain proteins in addition to recombinant versions of the Sac1p phosphoinositide-phosphatase indicated that PI(4)P was required on Golgi membranes for fusion with coat protein complex II (COPII) vesicles. PI(4)P inhibition did not prevent vesicle tethering but significantly reduced formation of soluble n-ethylmaleimide sensitive factor adaptor protein receptor (SNARE) complexes between vesicle and Golgi SNARE proteins. Moreover, semi-intact cell membranes containing elevated levels of the ER-Golgi SNARE proteins and Sly1p were less sensitive to PI(4)P inhibitors. Finally, in vivo analyses of a pik1 mutant strain showed that inhibition of PI(4)P synthesis blocked anterograde transport from the ER to early Golgi compartments. Together, the data presented here indicate that PI(4)P is required for the SNARE-dependent fusion stage of COPII vesicles with the Golgi complex. |
format | Text |
id | pubmed-3020917 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-30209172011-03-30 Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments Lorente-Rodríguez, Andrés Barlowe, Charles Mol Biol Cell Articles The role of specific membrane lipids in transport between endoplasmic reticulum (ER) and Golgi compartments is poorly understood. Using cell-free assays that measure stages in ER-to-Golgi transport, we screened a variety of enzyme inhibitors, lipid-modifying enzymes, and lipid ligands to investigate requirements in yeast. The pleckstrin homology (PH) domain of human Fapp1, which binds phosphatidylinositol-4-phosphate (PI(4)P) specifically, was a strong and specific inhibitor of anterograde transport. Analysis of wild type and mutant PH domain proteins in addition to recombinant versions of the Sac1p phosphoinositide-phosphatase indicated that PI(4)P was required on Golgi membranes for fusion with coat protein complex II (COPII) vesicles. PI(4)P inhibition did not prevent vesicle tethering but significantly reduced formation of soluble n-ethylmaleimide sensitive factor adaptor protein receptor (SNARE) complexes between vesicle and Golgi SNARE proteins. Moreover, semi-intact cell membranes containing elevated levels of the ER-Golgi SNARE proteins and Sly1p were less sensitive to PI(4)P inhibitors. Finally, in vivo analyses of a pik1 mutant strain showed that inhibition of PI(4)P synthesis blocked anterograde transport from the ER to early Golgi compartments. Together, the data presented here indicate that PI(4)P is required for the SNARE-dependent fusion stage of COPII vesicles with the Golgi complex. The American Society for Cell Biology 2011-01-15 /pmc/articles/PMC3020917/ /pubmed/21119004 http://dx.doi.org/10.1091/mbc.E10-04-0317 Text en © 2011 Lorente-Rodríguez et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0). “ASCB®,“ “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society of Cell Biology. |
spellingShingle | Articles Lorente-Rodríguez, Andrés Barlowe, Charles Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments |
title | Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments |
title_full | Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments |
title_fullStr | Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments |
title_full_unstemmed | Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments |
title_short | Requirement for Golgi-localized PI(4)P in fusion of COPII vesicles with Golgi compartments |
title_sort | requirement for golgi-localized pi(4)p in fusion of copii vesicles with golgi compartments |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3020917/ https://www.ncbi.nlm.nih.gov/pubmed/21119004 http://dx.doi.org/10.1091/mbc.E10-04-0317 |
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