.... , Fabian , L. , Hayes , M. , Polevoy , G. , Bazinet , C. and Brill , J. A. ( 2008 ). Depletion of plasma membrane PtdIns(4,5)P2 reveals essential roles for phosphoinositides in flagellar biogenesis . J. Cell Sci.   121 , 1076 - 1084 . 10.1242/jcs.024927 Wong , R...

... at the periphery of these structures. Based on their shape and size, we refer to them as NLIs. Scale bar: 200 nm. The graph shows that PtdIns(4,5)P 2 molecules and NM1 significantly colocalise at the distance of 25–75 nm ( n =21, PCCF>1, ** P ≤0.01). Fig. 1. Nucleoplasmic PtdIns(4,5)P2 appears in small foci...

... by the orange stars. This is well-supported by the presence of the different PIPKs, PtdIns(4,5) P 2 effectors and PtdIns(4,5) P 2 phosphatases at intracellular compartments that are shown here. Fig. 1. Widespread subcellular distribution of PtdIns(4,5)P2 throughout mammalian cells. Although...

... of 10 µM ionomycin. Scale bars: 10 µm. Fig. 1. The presence of PMCA proteins prevents the translocation of the PtdIns(4,5)P2 sensor during ionomycin treatments. The PtdIns(4,5)P2 sensor, PHPLCδ1–RFP was expressed alone or together with GFP–PMCA2wb in sh-HeLa cells; or together with GFP-PMCA4b-LA...

.... mss4 mutants have reduced levels of PtdIns(4,5)P2, an altered distribution of plasma membrane PtdIns(4,5)P2 and accumulate mannosyl-di-inositolphosphate-ceramide in vivo. (A) Quantification of PtdIns(4)P (PIP) and PtdIns(4,5)P2 (PIP2) levels in indicated strains. Cells were labeled with [33P]Pi...

... , 11157 – 11161 . 10.1073/pnas.90.23.11157 Mason   D. , Mallo   G. V. , Terebiznik   M. R. , Payrastre   B. , Finlay   B. B. , Brumell   J. H. , Rameh   L. , Grinstein   S. ( 2007 ). Alteration of epithelial structure and function associated with PtdIns(4,5)P2 degradation...

.... , Bittner   M. A. ( 2000 ). A pleckstrin homology domain specific for phosphatidylinositol 4, 5-bisphosphate (PtdIns-4,5-P2) and fused to green fluorescent protein identifies plasma membrane PtdIns-4,5-P2 as being important in exocytosis.   J. Biol. Chem.   275 , 17878 – 17885 . 10.1074/jbc.M000925200...

... VIL/WT treated with the PLC-γ1 inhibitor U73122 show a significant increase ( P <0.001, n  = 30) in the average number of filopodia assembled per cell compared to cells treated with the negative control (U73343). Fig. 5. PtdIns(4,5)P2 regulates villin self-association and villin-induced...

...) cells were transfected with the constructs shown and were labelled with [32P]orthophosphate for 2 hours after which phosphoinositides were extracted and the levels of PtdIns(4,5)P2 were analysed and quantified. A duplicate transfection was used to determine the expression levels of GFP-PIP5K (bottom...

... are in the cytosol-exposed loops of the FERM domain ( Pearson et al., 2000 ). Fig. 1. Drosophila Moe binds to PtdIns(4,5)P2 in a liposome co-sedimentation assay. (A) MoeWT and MoeKN association with phosphatidylcholine liposomes containing increasing concentrations of PtdIns(4,5)P2. The supernatant contains...

... of PtdIns(4)P (blue) and PtdIns(4,5)P2 (orange) showing that the carbachol-induced changes in the PtdIns(4)P levels are intact, whereas the PtdIns(4,5)P2 response is reduced after exposure to 50 μM cyclopiazonic acid (CPA) in a Ca2+-deficient medium containing 2 mM EGTA. (E) Mean ± s.e.m. for the effect...

... for RhoGDI, inducing the activation of Rho, which results in cell retraction. Fig. 2. PIP5K and PtdIns(4,5)P2 are involved in processes requiring actin organisation and focal-adhesion regulation. (A) Different small GTPases can regulate PIP5K activity, resulting in effects on the actin cytoskeleton...

... ). An emerging role for PtdIns(4,5)P2-mediated signalling in human disease. Trends Pharmacol. Sci. 26 , 654 -660. Harootunian, A. T., Kao, J. P., Paranjape, S. and Tsien, R. Y. ( 1991 ). Generation of calcium oscillations in fibroblasts by positive feedback between calcium and IP3. Science 251 , 75...

... the location of point mutations. (A) ENTHR65A/K78A, but not ENTHT107A, exhibits impaired binding to PtdIns(4,5)P2. PtdIns and PtdIns(4,5)P2 (PI and PI(4,5)P2, respectively) were spotted onto nitrocellulose membrane and then incubated with lysate from Dictyostelium cells expressing either wild-type or mutant...

... of the NGF-receptor starts the PtdIns(4,5) P 2 hydrolysis cycle again, and creates a balanced level of free profilin I, which may then contribute to neurite elongation. Where inactive, proteins are shown in grey lettering. Fig. 7. Model for the regulation of profilin-I-actin interaction by PtdIns(4,5)P2...

... to form PtdIns(4)P by type III PtdIns 4-kinase (step 3), which is inhibited by wortmannin (WT) and LY294002 (LY). PtdIns(4)P is phosphorylated by PtdIns(4)P 5-kinase to form phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2]. This in turn, is hydrolyzed by phospholipase C (PLC; step 4) to form Ins...