title: Type Igamma PIP kinase is a novel uropod component that regulates rear retraction during neutrophil chemotaxis.
abstract: Cell polarization is necessary for directed migration and leukocyte recruitment to inflamed tissues. Recent progress has been made in defining the molecular mechanisms that regulate chemoattractant-induced cell polarity during chemotaxis, including the contribution of phosphoinositide 3-kinase (PI3K)-dependent phosphatidylinositol (3,4,5)-trisphosphate [PtdIns(3,4,5)P(3)] synthesis at the leading edge. However, less is known about the molecular composition of the cell rear and how the uropod functions during cell motility. Here, we demonstrate that phosphatidylinositol phosphate kinase type Igamma (PIPKIgamma661), which generates PtdIns(4,5)P(2), is enriched in the uropod during chemotaxis of primary neutrophils and differentiated HL-60 cells (dHL-60). Using time-lapse microscopy, we show that enrichment of PIPKIgamma661 at the cell rear occurs early upon chemoattractant stimulation and is persistent during chemotaxis. Accordingly, we were able to detect enrichment of PtdIns(4,5)P(2) at the uropod during chemotaxis. Overexpression of kinase-dead PIPKIgamma661 compromised uropod formation and rear retraction similar to inhibition of ROCK signaling, suggesting that PtdIns(4,5)P(2) synthesis is important to elicit the backness response during chemotaxis. Together, our findings identify a previously unknown function for PIPKIgamma661 as a novel component of the backness signal that regulates rear retraction during chemotaxis.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Wisconsin, Madison, WI 53706, USA.
authors.forename: Mary A
authors.initials: MA
authors.lastname: Lokuta
authors.forename: Melissa A
authors.initials: MA
authors.lastname: Senetar
authors.forename: David A
authors.initials: DA
authors.lastname: Bennin
authors.forename: Paul A
authors.initials: PA
authors.lastname: Nuzzi
authors.forename: Keefe T
authors.initials: KT
authors.lastname: Chan
authors.forename: Vanessa L
authors.initials: VL
authors.lastname: Ott
authors.forename: Anna
authors.initials: A
authors.lastname: Huttenlocher
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D016764:Cell Polarity; D002633:Chemotaxis; D022001:Focal Adhesions / Q000378:metabolism; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D007962:Leukocytes / Q000166:cytology / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D009504:Neutrophils / Q000166:cytology* / Q000201:enzymology*; D018129:Phosphatidylinositol Phosphates / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D018129:Phosphatidylinositol Phosphates; C060974:phosphatidylinositol 3,4,5-triphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C020077:1-phosphatidylinositol-4-phosphate 5-kinase