Relationship Between Phosphatidylinositol 4-phosphate Synthesis, Membrane Organization, and Lateral Diffusion of PI4KIIalpha at the Trans-Golgi Network

Overview

Journal J Lipid Res

Publisher Elsevier

Specialty Biochemistry

Date 2010 Apr 15

PMID 20388919

Citations 30

Authors

Shane Minogue

K M Emily Chu

Emily J Westover

Douglas F Covey

J Justin Hsuan

Mark G Waugh

Affiliations

Soon will be listed here.

Abstract

Type II phosphatidylinositol 4-kinase IIalpha (PI4KIIalpha) is the dominant phosphatidylinositol kinase activity measured in mammalian cells and has important functions in intracellular vesicular trafficking. Recently PI4KIIalpha has been shown to have important roles in neuronal survival and tumorigenesis. This study focuses on the relationship between membrane cholesterol levels, phosphatidylinositol 4-phosphate (PI4P) synthesis, and PI4KIIalpha mobility. Enzyme kinetic measurements, sterol substitution studies, and membrane fragmentation analyses all revealed that cholesterol regulates PI4KIIalpha activity indirectly through effects on membrane structure. In particular, we found that cholesterol levels determined the distribution of PI4KIIalpha to biophysically distinct membrane domains. Imaging studies on cells expressing enhanced green fluorescent protein (eGFP)-tagged PI4KIIalpha demonstrated that cholesterol depletion resulted in morphological changes to the juxtanuclear membrane pool of the enzyme. Lateral membrane diffusion of eGFP-PI4KIIalpha was assessed by fluorescence recovery after photobleaching (FRAP) experiments, which revealed the existence of both mobile and immobile pools of the enzyme. Sterol depletion decreased the size of the mobile pool of PI4KIIalpha. Further measurements revealed that the reduction in the mobile fraction of PI4KIIalpha correlated with a loss of trans-Golgi network (TGN) membrane connectivity. We conclude that cholesterol modulates PI4P synthesis through effects on membrane organization and enzyme diffusion.

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References

Crowder C, Westover E, Kumar A, Ostlund Jr R, Covey D . Enantiospecificity of cholesterol function in vivo. J Biol Chem. 2001; 276(48):44369-72. DOI: 10.1074/jbc.C100535200. View

Meder D, Moreno M, Verkade P, Vaz W, Simons K . Phase coexistence and connectivity in the apical membrane of polarized epithelial cells. Proc Natl Acad Sci U S A. 2006; 103(2):329-34. PMC: 1324955. DOI: 10.1073/pnas.0509885103. View

van Meer G, Voelker D, Feigenson G . Membrane lipids: where they are and how they behave. Nat Rev Mol Cell Biol. 2008; 9(2):112-24. PMC: 2642958. DOI: 10.1038/nrm2330. View

Bock J, Klumperman J, Davanger S, Scheller R . Syntaxin 6 functions in trans-Golgi network vesicle trafficking. Mol Biol Cell. 1997; 8(7):1261-71. PMC: 276151. DOI: 10.1091/mbc.8.7.1261. View

Waugh M, Minogue S, Anderson J, Balinger A, Blumenkrantz D, Calnan D . Localization of a highly active pool of type II phosphatidylinositol 4-kinase in a p97/valosin-containing-protein-rich fraction of the endoplasmic reticulum. Biochem J. 2003; 373(Pt 1):57-63. PMC: 1223458. DOI: 10.1042/BJ20030089. View

Owen D, Williamson D, Rentero C, Gaus K . Quantitative microscopy: protein dynamics and membrane organisation. Traffic. 2009; 10(8):962-71. DOI: 10.1111/j.1600-0854.2009.00908.x. View

Simons J, Al-Shawi R, Minogue S, Waugh M, Wiedemann C, Evangelou S . Loss of phosphatidylinositol 4-kinase 2alpha activity causes late onset degeneration of spinal cord axons. Proc Natl Acad Sci U S A. 2009; 106(28):11535-9. PMC: 2710652. DOI: 10.1073/pnas.0903011106. View

Pike L . Lipid rafts: heterogeneity on the high seas. Biochem J. 2003; 378(Pt 2):281-92. PMC: 1223991. DOI: 10.1042/BJ20031672. View

Levine T, Munro S . Targeting of Golgi-specific pleckstrin homology domains involves both PtdIns 4-kinase-dependent and -independent components. Curr Biol. 2002; 12(9):695-704. DOI: 10.1016/s0960-9822(02)00779-0. View

10.

Craige B, Salazar G, Faundez V . Phosphatidylinositol-4-kinase type II alpha contains an AP-3-sorting motif and a kinase domain that are both required for endosome traffic. Mol Biol Cell. 2008; 19(4):1415-26. PMC: 2291421. DOI: 10.1091/mbc.e07-12-1239. View

11.

Orci L, Montesano R, Meda P, Malaisse-Lagae F, Brown D, Perrelet A . Heterogeneous distribution of filipin--cholesterol complexes across the cisternae of the Golgi apparatus. Proc Natl Acad Sci U S A. 1981; 78(1):293-7. PMC: 319039. DOI: 10.1073/pnas.78.1.293. View

12.

Kenworthy A, Nichols B, Remmert C, Hendrix G, Kumar M, Zimmerberg J . Dynamics of putative raft-associated proteins at the cell surface. J Cell Biol. 2004; 165(5):735-46. PMC: 2172371. DOI: 10.1083/jcb.200312170. View

13.

Barylko B, Mao Y, Wlodarski P, Jung G, Binns D, Sun H . Palmitoylation controls the catalytic activity and subcellular distribution of phosphatidylinositol 4-kinase II{alpha}. J Biol Chem. 2009; 284(15):9994-10003. PMC: 2665123. DOI: 10.1074/jbc.M900724200. View

14.

Banting G, Ponnambalam S . TGN38 and its orthologues: roles in post-TGN vesicle formation and maintenance of TGN morphology. Biochim Biophys Acta. 1997; 1355(3):209-17. DOI: 10.1016/s0167-4889(96)00146-2. View

15.

Pike L, Miller J . Cholesterol depletion delocalizes phosphatidylinositol bisphosphate and inhibits hormone-stimulated phosphatidylinositol turnover. J Biol Chem. 1998; 273(35):22298-304. DOI: 10.1074/jbc.273.35.22298. View

16.

Westover E, Covey D . The enantiomer of cholesterol. J Membr Biol. 2005; 202(2):61-72. DOI: 10.1007/s00232-004-0714-7. View

17.

Song K, Li Shengwen , Okamoto T, Quilliam L, Sargiacomo M, Lisanti M . Co-purification and direct interaction of Ras with caveolin, an integral membrane protein of caveolae microdomains. Detergent-free purification of caveolae microdomains. J Biol Chem. 1996; 271(16):9690-7. DOI: 10.1074/jbc.271.16.9690. View

18.

Stuven E, Porat A, Shimron F, Fass E, Kaloyanova D, Brugger B . Intra-Golgi protein transport depends on a cholesterol balance in the lipid membrane. J Biol Chem. 2003; 278(52):53112-22. DOI: 10.1074/jbc.M300402200. View

19.

Salazar G, Craige B, Wainer B, Guo J, De Camilli P, Faundez V . Phosphatidylinositol-4-kinase type II alpha is a component of adaptor protein-3-derived vesicles. Mol Biol Cell. 2005; 16(8):3692-704. PMC: 1182308. DOI: 10.1091/mbc.e05-01-0020. View

20.

Hao M, Lin S, Karylowski O, Wustner D, McGraw T, Maxfield F . Vesicular and non-vesicular sterol transport in living cells. The endocytic recycling compartment is a major sterol storage organelle. J Biol Chem. 2001; 277(1):609-17. DOI: 10.1074/jbc.M108861200. View