PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization

The lipid phosphatidylinositol-4,5-bisphosphate (PtdIns[4,5]P2) appears to play an important role in endocytosis. However, the timing of its formation and turnover, and its specific functions at different stages during endocytic internalization, have not been established. In this study, Sla2 ANTH-GF...

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Main Authors:	Sun, Yidi, Carroll, Susheela, Kaksonen, Marko, Toshima, Junko Y., Drubin, David G.
Format:	Online
Language:	English
Published:	The Rockefeller University Press 2007
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2064142/

id	pubmed-2064142
recordtype	oai_dc
spelling	pubmed-20641422007-11-29 PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization Sun, Yidi Carroll, Susheela Kaksonen, Marko Toshima, Junko Y. Drubin, David G. Research Articles The lipid phosphatidylinositol-4,5-bisphosphate (PtdIns[4,5]P2) appears to play an important role in endocytosis. However, the timing of its formation and turnover, and its specific functions at different stages during endocytic internalization, have not been established. In this study, Sla2 ANTH-GFP and Sjl2-3GFP were expressed as functional fusion proteins at endogenous levels to quantitatively explore PtdIns(4,5)P2 dynamics during endocytosis in yeast. Our results indicate that PtdIns(4,5)P2 levels increase and decline in conjunction with coat and actin assembly and disassembly, respectively. Live-cell image analysis of endocytic protein dynamics in an sjl1Δ sjl2Δ mutant, which has elevated PtdIns(4,5)P2 levels, revealed that the endocytic machinery is still able to assemble and disassemble dynamically, albeit nonproductively. The defects in the dynamic behavior of the various endocytic proteins in this double mutant suggest that PtdIns(4,5)P2 turnover is required for multiple stages during endocytic vesicle formation. Furthermore, our results indicate that PtdIns(4,5)P2 turnover may act in coordination with the Ark1/Prk1 protein kinases in stimulating disassembly of the endocytic machinery. The Rockefeller University Press 2007-04-23 /pmc/articles/PMC2064142/ /pubmed/17452534 http://dx.doi.org/10.1083/jcb.200611011 Text en Copyright © 2007, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Sun, Yidi Carroll, Susheela Kaksonen, Marko Toshima, Junko Y. Drubin, David G.
spellingShingle	Sun, Yidi Carroll, Susheela Kaksonen, Marko Toshima, Junko Y. Drubin, David G. PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization
author_facet	Sun, Yidi Carroll, Susheela Kaksonen, Marko Toshima, Junko Y. Drubin, David G.
author_sort	Sun, Yidi
title	PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization
title_short	PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization
title_full	PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization
title_fullStr	PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization
title_full_unstemmed	PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization
title_sort	ptdins(4,5)p2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization
description	The lipid phosphatidylinositol-4,5-bisphosphate (PtdIns[4,5]P2) appears to play an important role in endocytosis. However, the timing of its formation and turnover, and its specific functions at different stages during endocytic internalization, have not been established. In this study, Sla2 ANTH-GFP and Sjl2-3GFP were expressed as functional fusion proteins at endogenous levels to quantitatively explore PtdIns(4,5)P2 dynamics during endocytosis in yeast. Our results indicate that PtdIns(4,5)P2 levels increase and decline in conjunction with coat and actin assembly and disassembly, respectively. Live-cell image analysis of endocytic protein dynamics in an sjl1Δ sjl2Δ mutant, which has elevated PtdIns(4,5)P2 levels, revealed that the endocytic machinery is still able to assemble and disassemble dynamically, albeit nonproductively. The defects in the dynamic behavior of the various endocytic proteins in this double mutant suggest that PtdIns(4,5)P2 turnover is required for multiple stages during endocytic vesicle formation. Furthermore, our results indicate that PtdIns(4,5)P2 turnover may act in coordination with the Ark1/Prk1 protein kinases in stimulating disassembly of the endocytic machinery.
publisher	The Rockefeller University Press
publishDate	2007
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2064142/
_version_	1611406457976127488

PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization

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