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PMID: 18188447 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Loss of PIP5KIgamma, unlike other PIP5KI isoforms, impairs the integrity of the membrane cytoskeleton in murine megakaryocytes.

The Journal of clinical investigation ·Vol. 118 ·No. 2 ·2008-02-00 ·Pages 812-9

Wang Y, Litvinov RI, Chen X, Bach TL, Lian L, Petrich BG, Monkley SJ, Kanaho Y, Critchley DR, Sasaki T, Birnbaum MJ, Weisel JW, Hartwig J, Abrams CS

Abstract

Phosphatidylinositol-4,5-bisphosphate (PIP(2)) is an abundant phospholipid that contributes to second messenger formation and has also been shown to contribute to the regulation of cytoskeletal dynamics in all eukaryotic cells. Although the alpha, beta, and gamma isoforms of phosphatidylinositol-4-phosphate-5-kinase I (PIP5KI) all synthesize PIP2, mammalian cells usually contain more than one PIP5KI isoform. This raises the question of whether different isoforms of PIP5KI fulfill different functions. Given the speculated role of PIP(2) in platelet and megakaryocyte actin dynamics, we analyzed murine megakaryocytes lacking individual PIP5KI isoforms. PIP5KIgamma(-/-) megakaryocytes exhibited plasma membrane blebbing accompanied by a decreased association of the membrane with the cytoskeleton. This membrane defect was rescued by adding back wild-type PIP5KIgamma, but not by adding a catalytically inactive mutant or a splice variant lacking the talin-binding motif. Notably, both PIP5KIbeta- and PIP5KIgamma(-/-) cells had impaired PIP(2) synthesis. However, PIP5KIbeta-null cells lacked the membrane-cytoskeleton defect. Furthermore, overexpressing PIP5KIbeta in PIP5KIgamma(-/-) cells failed to revert this defect. Megakaryocytes lacking the PIP5KIgamma-binding partner, talin1, mimicked the membrane-cytoskeleton defect phenotype seen in PIP5KIgamma(-/-) cells. These findings demonstrate a unique role for PIP5KIgamma in the anchoring of the cell membrane to the cytoskeleton in megakaryocytes, probably through a pathway involving talin. These observations further demonstrate that individual PIP5KI isoforms fulfill distinct functions within cells.

MeSH Terms
Animals Cell Membrane/ultrastructure Cytoskeleton/enzymology,ultrastructure Male Megakaryocytes/enzymology,ultrastructure Mice Mice, Mutant Strains Phosphatidylinositol 4,5-Diphosphate/metabolism Phosphotransferases (Alcohol Group Acceptor)/genetics,metabolism Protein Isoforms/genetics,metabolism Talin/metabolism
Chemicals
Phosphatidylinositol 4,5-Diphosphate Protein Isoforms Talin Phosphotransferases (Alcohol Group Acceptor) 1-phosphatidylinositol-4-phosphate 5-kinase
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Wang Yanfeng
Department of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Litvinov Rustem I
Chen Xinsheng
Bach Tami L
Lian Lurong
Petrich Brian G
Monkley Susan J
Kanaho Yasunori
Critchley David R
Sasaki Takehiko
Birnbaum Morris J
Weisel John W
Hartwig John
Abrams Charles S
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Article Info
Journal
The Journal of clinical investigation
Abbr.
J Clin Invest
ISSN
0021-9738
DOI
10.1172/JCI34239
Published
2008-02-00
Pages
812-9
Language
English
Region
United States
NLM ID
7802877
PMCID
PMC2176194
Subset
IM
Grants
Wellcome Trust · United Kingdom
Corrections
ErratumIn
-
External Links
PubMed source DOI: 10.1172/JCI34239 PMC Full Text View journal details
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