Home LiteratureArticle Details
PMID: 24711504 Published · ppublish English Journal Article Research Support, N.I.H., Intramural Video-Audio Media

A novel probe for phosphatidylinositol 4-phosphate reveals multiple pools beyond the Golgi.

The Journal of cell biology ·Vol. 205 ·No. 1 ·2014-04-14 ·Pages 113-26

Hammond GR, Machner MP, Balla T

Abstract

Polyphosphoinositides are an important class of lipid that recruit specific effector proteins to organelle membranes. One member, phosphatidylinositol 4-phosphate (PtdIns4P) has been localized to Golgi membranes based on the distribution of lipid binding modules from PtdIns4P effector proteins. However, these probes may be biased by additional interactions with other Golgi-specific determinants. In this paper, we derive a new PtdIns4P biosensor using the PtdIns4P binding of SidM (P4M) domain of the secreted effector protein SidM from the bacterial pathogen Legionella pneumophila. PtdIns4P was necessary and sufficient for localization of P4M, which revealed pools of the lipid associated not only with the Golgi but also with the plasma membrane and Rab7-positive late endosomes/lysosomes. PtdIns4P distribution was determined by the localization and activities of both its anabolic and catabolic enzymes. Therefore, P4M reports a wider cellular distribution of PtdIns4P than previous probes and therefore will be valuable for dissecting the biological functions of PtdIns4P in its assorted membrane compartments.

MeSH Terms
Animals Bacterial Proteins/genetics,metabolism Binding Sites Biosensing Techniques COS Cells Cell Membrane/metabolism Chlorocebus aethiops Dogs Endosomes/metabolism Genes, Reporter Golgi Apparatus/metabolism Guanine Nucleotide Exchange Factors/genetics,metabolism Luminescent Proteins/genetics,metabolism Lysosomes/metabolism Mice Microscopy, Confocal Minor Histocompatibility Antigens Phosphatidylinositol Phosphates/metabolism Phosphotransferases (Alcohol Group Acceptor)/metabolism Protein Structure, Tertiary Recombinant Fusion Proteins/metabolism Second Messenger Systems Time Factors Time-Lapse Imaging Transfection rab GTP-Binding Proteins/metabolism rab7 GTP-Binding Proteins
Chemicals
Bacterial Proteins Guanine Nucleotide Exchange Factors Luminescent Proteins Minor Histocompatibility Antigens Phosphatidylinositol Phosphates Recombinant Fusion Proteins SidM protein, Legionella pneumophila phosphatidylinositol 4-phosphate rab7 GTP-Binding Proteins rab7 GTP-binding proteins, human rab7 GTP-binding proteins, mouse Phosphotransferases (Alcohol Group Acceptor) phosphatidylinositol phosphate 4-kinase rab GTP-Binding Proteins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hammond Gerald R V
Program in Developmental Neuroscience and 2 Cell Biology and Metabolism Program, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
Machner Matthias P
Balla Tamas
References (82)
82 references, click to expand
  1. Phylogenomics of phosphoinositide lipid kinases: perspectives on the evolution of second messenger signaling and drug discovery.
    BMC Evol Biol. 2011 Jan 05;11:4 PMID: 21208444
  2. Phosphatidylinositol-4-kinase type II alpha is a component of adaptor protein-3-derived vesicles.
    Mol Biol Cell. 2005 Aug;16(8):3692-704 PMID: 15944223
  3. Inositol lipid regulation of lipid transfer in specialized membrane domains.
    Trends Cell Biol. 2013 Jun;23(6):270-8 PMID: 23489878
  4. Direct involvement of phosphatidylinositol 4-phosphate in secretion in the yeast Saccharomyces cerevisiae.
    J Biol Chem. 1999 Nov 26;274(48):34294-300 PMID: 10567405
  5. Membrane phosphatidylserine regulates surface charge and protein localization.
    Science. 2008 Jan 11;319(5860):210-3 PMID: 18187657
  6. FAPPs control Golgi-to-cell-surface membrane traffic by binding to ARF and PtdIns(4)P.
    Nat Cell Biol. 2004 May;6(5):393-404 PMID: 15107860
  7. Rapid redistribution of Golgi proteins into the ER in cells treated with brefeldin A: evidence for membrane cycling from Golgi to ER.
    Cell. 1989 Mar 10;56(5):801-13 PMID: 2647301
  8. Subcellular localization of phosphatidylinositol 4,5-bisphosphate using the pleckstrin homology domain of phospholipase C delta1.
    Biochem J. 2002 May 1;363(Pt 3):657-66 PMID: 11964166
  9. Manipulation of host membranes by bacterial effectors.
    Nat Rev Microbiol. 2011 Jul 18;9(9):635-46 PMID: 21765451
  10. Integration of Golgi trafficking and growth factor signaling by the lipid phosphatase SAC1.
    J Cell Biol. 2008 Feb 25;180(4):803-12 PMID: 18299350
  11. Organelle-specific, rapid induction of molecular activities and membrane tethering.
    Nat Methods. 2010 Mar;7(3):206-8 PMID: 20154678
  12. PI4P and PI(4,5)P2 are essential but independent lipid determinants of membrane identity.
    Science. 2012 Aug 10;337(6095):727-30 PMID: 22722250
  13. High-affinity binding of phosphatidylinositol 4-phosphate by Legionella pneumophila DrrA.
    EMBO Rep. 2010 Aug;11(8):598-604 PMID: 20616805
  14. Visualization of phosphoinositides that bind pleckstrin homology domains: calcium- and agonist-induced dynamic changes and relationship to myo-[3H]inositol-labeled phosphoinositide pools.
    J Cell Biol. 1998 Oct 19;143(2):501-10 PMID: 9786958
  15. Cholesterol sensor ORP1L contacts the ER protein VAP to control Rab7-RILP-p150 Glued and late endosome positioning.
    J Cell Biol. 2009 Jun 29;185(7):1209-25 PMID: 19564404
  16. Rapid chemically induced changes of PtdIns(4,5)P2 gate KCNQ ion channels.
    Science. 2006 Dec 1;314(5804):1454-7 PMID: 16990515
  17. SAC1-like domains of yeast SAC1, INP52, and INP53 and of human synaptojanin encode polyphosphoinositide phosphatases.
    J Biol Chem. 1999 May 7;274(19):12990-5 PMID: 10224048
  18. Maintenance of hormone-sensitive phosphoinositide pools in the plasma membrane requires phosphatidylinositol 4-kinase IIIalpha.
    Mol Biol Cell. 2008 Feb;19(2):711-21 PMID: 18077555
  19. A new class of carriers that transport selective cargo from the trans Golgi network to the cell surface.
    EMBO J. 2012 Oct 17;31(20):3976-90 PMID: 22909819
  20. The rab7 GTPase resides on a vesicular compartment connected to lysosomes.
    J Cell Sci. 1995 Nov;108 ( Pt 11):3349-58 PMID: 8586647
  21. Perturbation of the morphology of the trans-Golgi network following Brefeldin A treatment: redistribution of a TGN-specific integral membrane protein, TGN38.
    J Cell Biol. 1992 Jan;116(1):85-94 PMID: 1730751
  22. Clathrin and phosphatidylinositol-4,5-bisphosphate regulate autophagic lysosome reformation.
    Nat Cell Biol. 2012 Sep;14(9):924-34 PMID: 22885770
  23. The yeast phosphatidylinositol-4-OH kinase pik1 regulates secretion at the Golgi.
    Nat Cell Biol. 1999 Dec;1(8):523-5 PMID: 10587649
  24. Phosphatidylinositol 4-kinase IIIbeta regulates the transport of ceramide between the endoplasmic reticulum and Golgi.
    J Biol Chem. 2006 Nov 24;281(47):36369-77 PMID: 17003043
  25. PI(3,4,5)P3 and PI(4,5)P2 lipids target proteins with polybasic clusters to the plasma membrane.
    Science. 2006 Dec 1;314(5804):1458-61 PMID: 17095657
  26. Coincidence detection in phosphoinositide signaling.
    Trends Cell Biol. 2005 Oct;15(10):540-7 PMID: 16139503
  27. Structural mechanism of host Rab1 activation by the bifunctional Legionella type IV effector SidM/DrrA.
    Proc Natl Acad Sci U S A. 2010 Mar 9;107(10):4699-704 PMID: 20176951
  28. Distinct Golgi populations of phosphatidylinositol 4-phosphate regulated by phosphatidylinositol 4-kinases.
    J Biol Chem. 2005 Mar 18;280(11):10501-8 PMID: 15634669
  29. Recruitment and activation of a lipid kinase by hepatitis C virus NS5A is essential for integrity of the membranous replication compartment.
    Cell Host Microbe. 2011 Jan 20;9(1):32-45 PMID: 21238945
  30. Impaired α-TTP-PIPs interaction underlies familial vitamin E deficiency.
    Science. 2013 May 31;340(6136):1106-10 PMID: 23599266
  31. Rab1 guanine nucleotide exchange factor SidM is a major phosphatidylinositol 4-phosphate-binding effector protein of Legionella pneumophila.
    J Biol Chem. 2009 Feb 20;284(8):4846-56 PMID: 19095644
  32. Fiji: an open-source platform for biological-image analysis.
    Nat Methods. 2012 Jun 28;9(7):676-82 PMID: 22743772
  33. Multiple pools of phosphatidylinositol 4-phosphate detected using the pleckstrin homology domain of Osh2p.
    J Biol Chem. 2004 Oct 22;279(43):44683-9 PMID: 15271978
  34. Coordination of Golgi functions by phosphatidylinositol 4-kinases.
    Trends Cell Biol. 2011 Feb;21(2):113-21 PMID: 21282087
  35. Assembly of the PtdIns 4-kinase Stt4 complex at the plasma membrane requires Ypp1 and Efr3.
    J Cell Biol. 2008 Dec 15;183(6):1061-74 PMID: 19075114
  36. A plasma membrane pool of phosphatidylinositol 4-phosphate is generated by phosphatidylinositol 4-kinase type-III alpha: studies with the PH domains of the oxysterol binding protein and FAPP1.
    Mol Biol Cell. 2005 Mar;16(3):1282-95 PMID: 15635101
  37. The lipid kinase phosphatidylinositol-4 kinase III alpha regulates the phosphorylation status of hepatitis C virus NS5A.
    PLoS Pathog. 2013 May;9(5):e1003359 PMID: 23675303
  38. Phosphoinositides: tiny lipids with giant impact on cell regulation.
    Physiol Rev. 2013 Jul;93(3):1019-137 PMID: 23899561
  39. Phosphatidylinositol-5-phosphate activation and conserved substrate specificity of the myotubularin phosphatidylinositol 3-phosphatases.
    Curr Biol. 2003 Mar 18;13(6):504-9 PMID: 12646134
  40. Phosphatidylinositol 4-kinase is required for endosomal trafficking and degradation of the EGF receptor.
    J Cell Sci. 2006 Feb 1;119(Pt 3):571-81 PMID: 16443754
  41. Hepatitis C virus stimulates the phosphatidylinositol 4-kinase III alpha-dependent phosphatidylinositol 4-phosphate production that is essential for its replication.
    J Virol. 2011 Sep;85(17):8870-83 PMID: 21697487
  42. Bright and stable near-infrared fluorescent protein for in vivo imaging.
    Nat Biotechnol. 2011 Jul 17;29(8):757-61 PMID: 21765402
  43. GOLPH3 bridges phosphatidylinositol-4- phosphate and actomyosin to stretch and shape the Golgi to promote budding.
    Cell. 2009 Oct 16;139(2):337-51 PMID: 19837035
  44. Phosphatidylinositol 4 phosphate regulates targeting of clathrin adaptor AP-1 complexes to the Golgi.
    Cell. 2003 Aug 8;114(3):299-310 PMID: 12914695
  45. A temporal threshold for formaldehyde crosslinking and fixation.
    PLoS One. 2009;4(2):e4636 PMID: 19247482
  46. Genetically encoded fluorescent probe to visualize intracellular phosphatidylinositol 3,5-bisphosphate localization and dynamics.
    Proc Natl Acad Sci U S A. 2013 Dec 24;110(52):21165-70 PMID: 24324172
  47. Rapidly inducible changes in phosphatidylinositol 4,5-bisphosphate levels influence multiple regulatory functions of the lipid in intact living cells.
    J Cell Biol. 2006 Nov 6;175(3):377-82 PMID: 17088424
  48. PI4KIIIα is required for cortical integrity and cell polarity during Drosophila oogenesis.
    J Cell Sci. 2014 Mar 1;127(Pt 5):954-66 PMID: 24413170
  49. Regulation of retromer recruitment to endosomes by sequential action of Rab5 and Rab7.
    J Cell Biol. 2008 Nov 3;183(3):513-26 PMID: 18981234
  50. How accurately can we image inositol lipids in living cells?
    Trends Pharmacol Sci. 2000 Jul;21(7):238-41 PMID: 10871889
  51. Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites.
    Cell. 2011 Feb 4;144(3):389-401 PMID: 21295699
  52. Localization of phosphatidylinositol 3-phosphate in yeast and mammalian cells.
    EMBO J. 2000 Sep 1;19(17):4577-88 PMID: 10970851
  53. Relationship between phosphatidylinositol 4-phosphate synthesis, membrane organization, and lateral diffusion of PI4KIIalpha at the trans-Golgi network.
    J Lipid Res. 2010 Aug;51(8):2314-24 PMID: 20388919
  54. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein.
    Nat Biotechnol. 2004 Dec;22(12):1567-72 PMID: 15558047
  55. Design of drug-resistant alleles of type-III phosphatidylinositol 4-kinases using mutagenesis and molecular modeling.
    Biochemistry. 2008 Feb 12;47(6):1599-607 PMID: 18205404
  56. Phosphoinositides in constitutive membrane traffic.
    Physiol Rev. 2004 Jul;84(3):699-730 PMID: 15269334
  57. Induction of a unique gene expression profile in primary human hepatocytes by hepatitis C virus core, NS3 and NS5A proteins.
    Carcinogenesis. 2007 Jul;28(7):1552-60 PMID: 17404395
  58. Immunocytochemical techniques reveal multiple, distinct cellular pools of PtdIns4P and PtdIns(4,5)P(2).
    Biochem J. 2009 Jul 29;422(1):23-35 PMID: 19508231
  59. The lipid kinase PI4KIIIβ preserves lysosomal identity.
    EMBO J. 2013 Feb 6;32(3):324-39 PMID: 23258225
  60. A bifunctional bacterial protein links GDI displacement to Rab1 activation.
    Science. 2007 Nov 9;318(5852):974-7 PMID: 17947549
  61. A four-step cycle driven by PI(4)P hydrolysis directs sterol/PI(4)P exchange by the ER-Golgi tether OSBP.
    Cell. 2013 Nov 7;155(4):830-43 PMID: 24209621
  62. A monomeric red fluorescent protein.
    Proc Natl Acad Sci U S A. 2002 Jun 11;99(12):7877-82 PMID: 12060735
  63. Compartmental signal modulation: Endosomal phosphatidylinositol 3-phosphate controls endosome morphology and selective cargo sorting.
    Proc Natl Acad Sci U S A. 2006 Oct 17;103(42):15473-8 PMID: 17030795
  64. Characterization of type II phosphatidylinositol 4-kinase isoforms reveals association of the enzymes with endosomal vesicular compartments.
    J Biol Chem. 2002 May 31;277(22):20041-50 PMID: 11923287
  65. Dual roles for the Drosophila PI 4-kinase four wheel drive in localizing Rab11 during cytokinesis.
    J Cell Biol. 2009 Dec 14;187(6):847-58 PMID: 19995935
  66. Optogenetic control of phosphoinositide metabolism.
    Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):E2316-23 PMID: 22847441
  67. Acute manipulation of Golgi phosphoinositides to assess their importance in cellular trafficking and signaling.
    Proc Natl Acad Sci U S A. 2010 May 4;107(18):8225-30 PMID: 20404150
  68. ARF mediates recruitment of PtdIns-4-OH kinase-beta and stimulates synthesis of PtdIns(4,5)P2 on the Golgi complex.
    Nat Cell Biol. 1999 Sep;1(5):280-7 PMID: 10559940
  69. The small GTPase rab5 functions as a regulatory factor in the early endocytic pathway.
    Cell. 1992 Sep 4;70(5):715-28 PMID: 1516130
  70. A novel HPLC-based approach makes possible the spatial characterization of cellular PtdIns5P and other phosphoinositides.
    Biochem J. 2010 May 27;428(3):375-84 PMID: 20370717
  71. An automated method to quantify and visualize colocalized fluorescent signals.
    J Neurosci Methods. 2005 Jul 15;146(1):42-9 PMID: 15935219
  72. Two phosphatidylinositol 4-kinases control lysosomal delivery of the Gaucher disease enzyme, β-glucocerebrosidase.
    Mol Biol Cell. 2012 Apr;23(8):1533-45 PMID: 22337770
  73. Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%.
    Nat Commun. 2012 Mar 20;3:751 PMID: 22434194
  74. Targeting of Golgi-specific pleckstrin homology domains involves both PtdIns 4-kinase-dependent and -independent components.
    Curr Biol. 2002 Apr 30;12(9):695-704 PMID: 12007412
  75. Functional characterization of a mammalian Sac1 and mutants exhibiting substrate-specific defects in phosphoinositide phosphatase activity.
    J Biol Chem. 2000 Nov 3;275(44):34293-305 PMID: 10887188
  76. Type II phosphatidylinositol 4-kinase regulates trafficking of secretory granule proteins in Drosophila.
    Development. 2012 Aug;139(16):3040-50 PMID: 22791894
  77. The human phosphatidylinositol phosphatase SAC1 interacts with the coatomer I complex.
    J Biol Chem. 2003 Dec 26;278(52):52689-99 PMID: 14527956
  78. Ci-VSP is a depolarization-activated phosphatidylinositol-4,5-bisphosphate and phosphatidylinositol-3,4,5-trisphosphate 5'-phosphatase.
    J Biol Chem. 2009 Jan 23;284(4):2106-13 PMID: 19047057
  79. A mammalian protein targeted by G1-arresting rapamycin-receptor complex.
    Nature. 1994 Jun 30;369(6483):756-8 PMID: 8008069
  80. PtdIns4P synthesis by PI4KIIIα at the plasma membrane and its impact on plasma membrane identity.
    J Cell Biol. 2012 Dec 10;199(6):1003-16 PMID: 23229899
  81. Glycosphingolipid synthesis requires FAPP2 transfer of glucosylceramide.
    Nature. 2007 Sep 6;449(7158):62-7 PMID: 17687330
  82. The Legionella effector protein DrrA AMPylates the membrane traffic regulator Rab1b.
    Science. 2010 Aug 20;329(5994):946-9 PMID: 20651120
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
1540-8140
DOI
10.1083/jcb.201312072
Published
2014-04-14
Epub
2014-00-07
Pages
113-26
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC3987136
Subset
IM
Grants
Intramural NIH HHS · United States
External Links
PubMed source DOI: 10.1083/jcb.201312072 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2