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PMID: 18843041 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Ca2+-regulated pool of phosphatidylinositol-3-phosphate produced by phosphatidylinositol 3-kinase C2alpha on neurosecretory vesicles.

Molecular biology of the cell ·Vol. 19 ·No. 12 ·2008-12-00 ·Pages 5593-603

Wen PJ, Osborne SL, Morrow IC, Parton RG, Domin J, Meunier FA

Abstract

Phosphatidylinositol-3-phosphate [PtdIns(3)P] is a key player in early endosomal trafficking and is mainly produced by class III phosphatidylinositol 3-kinase (PI3K). In neurosecretory cells, class II PI3K-C2alpha and its lipid product PtdIns(3)P have recently been shown to play a critical role during neuroexocytosis, suggesting that two distinct pools of PtdIns(3)P might coexist in these cells. However, the precise characterization of this additional pool of PtdIns(3)P remains to be established. Using a selective PtdIns(3)P probe, we have identified a novel PtdIns(3)P-positive pool localized on secretory vesicles, sensitive to PI3K-C2alpha knockdown and relatively resistant to wortmannin treatment. In neurosecretory cells, stimulation of exocytosis promoted a transient albeit large increase in PtdIns(3)P production localized on secretory vesicles sensitive to PI3K-C2alpha knockdown and expression of PI3K-C2alpha catalytically inactive mutant. Using purified chromaffin granules, we found that PtdIns(3)P production is controlled by Ca(2+). We confirmed that PtdIns(3)P production from recombinantly expressed PI3K-C2alpha is indeed regulated by Ca(2+). We provide evidence that a dynamic pool of PtdIns(3)P synthesized by PI3K-C2alpha occurs on secretory vesicles in neurosecretory cells, demonstrating that the activity of a member of the PI3K family is regulated by Ca(2+) in vitro and in living neurosecretory cells.

MeSH Terms
Androstadienes/metabolism Animals Calcium/metabolism Cattle Cell Line Chromaffin Cells/cytology,metabolism Class II Phosphatidylinositol 3-Kinases Exocytosis/physiology Humans Phosphatidylinositol 3-Kinases/genetics,metabolism Phosphatidylinositol Phosphates/metabolism Protein Kinase Inhibitors/metabolism RNA, Small Interfering/genetics,metabolism Rats Recombinant Fusion Proteins/genetics,metabolism Secretory Vesicles/metabolism Signal Transduction/physiology Wortmannin
Chemicals
Androstadienes Phosphatidylinositol Phosphates Protein Kinase Inhibitors RNA, Small Interfering Recombinant Fusion Proteins phosphatidylinositol 3-phosphate Phosphatidylinositol 3-Kinases Class II Phosphatidylinositol 3-Kinases Calcium Wortmannin
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Wen Peter J
Molecular Dynamics of Synaptic Function Laboratory, Queensland Brain Institute and School of Biomedical Sciences, The University of Queensland, St. Lucia, 4072 Queensland, Australia.
Osborne Shona L
Morrow Isabel C
Parton Robert G
Domin Jan
Meunier Frederic A
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
DOI
10.1091/mbc.e08-06-0595
Published
2008-12-00
Epub
2008-00-08
Pages
5593-603
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC2592681
Subset
IM
Grants
Medical Research Council · G0500936 · United Kingdom
External Links
PubMed source DOI: 10.1091/mbc.e08-06-0595 PMC Full Text View journal details
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