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PMID: 15809313 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Phosphatidylinositol-4,5-bisphosphate hydrolysis directs actin remodeling during phagocytosis.

The Journal of cell biology ·Vol. 169 ·No. 1 ·2005-04-11 ·Pages 139-49

Scott CC, Dobson W, Botelho RJ, Coady-Osberg N, Chavrier P, Knecht DA, Heath C, Stahl P, Grinstein S

Abstract

The Rho GTPases play a critical role in initiating actin polymerization during phagocytosis. In contrast, the factors directing the disassembly of F-actin required for fission of the phagocytic vacuole are ill defined. We used fluorescent chimeric proteins to monitor the dynamics of association of actin and active Cdc42 and Rac1 with the forming phagosome. Although actin was found to disappear from the base of the forming phagosome before sealing was complete, Rac1/Cdc42 activity persisted, suggesting that termination of GTPase activity is not the main determinant of actin disassembly. Furthermore, fully internalized phagosomes engineered to associate constitutively with active Rac1 showed little associated F-actin. The disappearance of phosphatidylinositol-4,5-bisphosphate (PI(4,5)P(2)) from the phagosomal membrane closely paralleled the course of actin disassembly. Furthermore, inhibition of PI(4,5)P(2) hydrolysis or increased PI(4,5)P(2) generation by overexpression of phosphatidylinositol phosphate kinase I prevented the actin disassembly necessary for the completion of phagocytosis. These observations suggest that hydrolysis of PI(4,5)P(2) dictates the remodeling of actin necessary for completion of phagocytosis.

MeSH Terms
Actins/chemistry,metabolism Animals Cell Line Dimerization Erythrocytes/cytology Green Fluorescent Proteins Humans Hydrolysis Mice Phagocytosis/physiology Phagosomes/metabolism Phosphatidylinositol 4,5-Diphosphate/metabolism Phosphotransferases (Alcohol Group Acceptor)/genetics,metabolism Recombinant Proteins/genetics,metabolism cdc42 GTP-Binding Protein/genetics,metabolism rac1 GTP-Binding Protein/genetics,metabolism
Chemicals
Actins Phosphatidylinositol 4,5-Diphosphate Recombinant Proteins Green Fluorescent Proteins Phosphotransferases (Alcohol Group Acceptor) cdc42 GTP-Binding Protein rac1 GTP-Binding Protein
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Scott Cameron C
Division of Cell Biology, The Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada.
Dobson Wendy
Botelho Roberto J
Coady-Osberg Natasha
Chavrier Philippe
Knecht David A
Heath Colin
Stahl Philip
Grinstein Sergio
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.200412162
Published
2005-04-11
Epub
2005-00-04
Pages
139-49
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2171893
Subset
IM
Grants
NIGMS NIH HHS · GM30599 · United States
External Links
PubMed source DOI: 10.1083/jcb.200412162 PMC Full Text View journal details
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