title: Localization of phosphatidylinositol 4,5-bisphosphate to lipid rafts and uroids in the human protozoan parasite Entamoeba histolytica.
abstract: Entamoeba histolytica is an intestinal protozoan parasite and is the causative agent of amoebiasis. During invasive infection, highly motile amoebae destroy the colonic epithelium, enter the blood circulation, and disseminate to other organs such as liver, causing liver abscess. Motility is a key factor in E. histolytica pathogenesis, and this process relies on a dynamic actomyosin cytoskeleton. In other systems, phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] is known to regulate a wide variety of cellular functions, including signal transduction, actin remodeling, and cell motility. Little is known about the role of PI(4,5)P2 in E. histolytica pathogenicity. In this study, we demonstrate that PI(4,5)P2 is localized to cholesterol-rich microdomains, lipid rafts, and the actin-rich fractions of the E. histolytica membrane. Microscopy revealed that the trailing edge of polarized trophozoites, uroids, are highly enriched in lipid rafts and their constituent lipid, PI(4,5)P2. Polarization and enrichment of uroids and rafts with PI(4,5)P2 were enhanced upon treatment of E. histolytica cells with cholesterol. Exposure to cholesterol also increased intracellular calcium, which is a downstream effector of PI(4,5)P2, with a concomitant increase in motility. Together, our data suggest that in E. histolytica, PI(4,5)P2 may signal from lipid rafts and cholesterol may play a role in triggering PI(4,5)P2-mediated signaling to enhance the motility of this pathogen.
authors.affiliation: Department of Genetics and Biochemistry, Clemson University, Clemson, South Carolina, USA.
authors.forename: Amrita B
authors.initials: AB
authors.lastname: Koushik
authors.forename: Rhonda R
authors.initials: RR
authors.lastname: Powell
authors.forename: Lesly A
authors.initials: LA
authors.lastname: Temesvari
mesh_terms: D001692:Biological Transport; D002465:Cell Movement / Q000502:physiology; D004748:Entamoeba histolytica / Q000166:cytology* / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism*; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D019269:Phosphatidylinositol 4,5-Diphosphate
references.citation: J Biol Chem. 2008 Oct 31;283(44):29920-8
references.pmid: 18723502
references.citation: Cell Calcium. 2009 Jun;45(6):554-65
references.pmid: 19376575
references.citation: Cell. 2000 Jan 21;100(2):221-8
references.pmid: 10660045
references.citation: Biochem Biophys Res Commun. 1998 Apr 28;245(3):684-90
references.pmid: 9588175
references.citation: Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18751-6
references.pmid: 17132728
references.citation: Arch Med Res. 2000 Jul-Aug;31(4 Suppl):S140-2
references.pmid: 11070257
references.citation: Mol Microbiol. 1996 Dec;22(5):849-57
references.pmid: 8971707
references.citation: Exp Parasitol. 2006 Apr;112(4):221-31
references.pmid: 16387299
references.citation: Trends Microbiol. 1996 May;4(5):191-7
references.pmid: 8727599
references.citation: J Cell Biol. 2004 Oct 25;167(2):327-37
references.pmid: 15504914
references.citation: Nat Rev Mol Cell Biol. 2009 May;10(5):353-9
references.pmid: 19373240
references.citation: Biochem J. 2002 May 1;363(Pt 3):657-66
references.pmid: 11964166
references.citation: Mol Biochem Parasitol. 2004 May;135(1):31-8
references.pmid: 15287584
references.citation: Blood. 2002 Feb 1;99(3):978-84
references.pmid: 11807002
references.citation: Dev Cell. 2002 Oct;3(4):469-78
references.pmid: 12408799
references.citation: PLoS Negl Trop Dis. 2011 Apr 05;5(4):e1002
references.pmid: 21483708
references.citation: Cell Signal. 2001 Jun;13(6):377-87
references.pmid: 11384836
references.citation: Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9642-7
references.pmid: 11493690
references.citation: Nature. 1995 Sep 7;377(6544):75-9
references.pmid: 7544874
references.citation: Biochem J. 1984 Jun 1;220(2):345-60
references.pmid: 6146314
references.citation: Exp Parasitol. 2008 Oct;120(2):127-34
references.pmid: 18588878
references.citation: Mol Biol Cell. 1996 Jun;7(6):843-51
references.pmid: 8816992
references.citation: Infect Immun. 2004 Sep;72(9):5349-57
references.pmid: 15322032
references.citation: J Exp Med. 1980 Aug 1;152(2):391-404
references.pmid: 6249883
references.citation: Biochemistry. 2009 Sep 1;48(34):8241-8
references.pmid: 19630438
references.citation: Microbes Infect. 2000 May;2(6):643-9
references.pmid: 10884615
references.citation: IUBMB Life. 2001 Jun;51(6):337-44
references.pmid: 11758800
references.citation: Mol Biochem Parasitol. 2002 May;121(2):254-64
references.pmid: 12034459
references.citation: Sci STKE. 2001 Dec 04;2001(111):re19
references.pmid: 11734659
references.citation: Annu Rev Biophys Biomol Struct. 2000;29:49-79
references.pmid: 10940243
references.citation: Lipids Health Dis. 2005 May 09;4:10
references.pmid: 15882457
references.citation: Traffic. 2003 Nov;4(11):724-38
references.pmid: 14617356
references.citation: Immunol Endocr Metab Agents Med Chem. 2008 Dec 1;8(4):349-357
references.pmid: 19956793
references.citation: Exp Cell Res. 2009 Apr 1;315(6):981-95
references.pmid: 19331818
references.citation: J Biol Chem. 2001 May 25;276(21):18640-8
references.pmid: 11278598
references.citation: Nat Immunol. 2008 Jul;9(7):743-52
references.pmid: 18536720
references.citation: Can J Microbiol. 2010 Dec;56(12):987-95
references.pmid: 21164568
references.citation: Nature. 1984 Nov 22-28;312(5992):315-21
references.pmid: 6095092
references.citation: J Cell Biol. 2007 Dec 31;179(7):1539-53
references.pmid: 18158329
references.citation: Infect Immun. 2005 Mar;73(3):1771-8
references.pmid: 15731078
references.citation: Int J Parasitol. 2011 Nov;41(13-14):1409-19
references.pmid: 22085647
references.citation: Infect Immun. 2010 Jan;78(1):125-37
references.pmid: 19901063
references.citation: Eukaryot Cell. 2012 Jun;11(6):743-51
references.pmid: 22505337
references.citation: Immunity. 2003 Nov;19(5):669-78
references.pmid: 14614854
references.citation: J Cell Sci. 1999 Apr;112 ( Pt 8):1191-201
references.pmid: 10085254
references.citation: Microbiology (Reading). 2004 Aug;150(Pt 8):2513-2525
references.pmid: 15289548
references.citation: Exp Parasitol. 1996 Jan;82(1):11-20
references.pmid: 8617326
references.citation: Mol Biol Cell. 2005 Dec;16(12):5773-83
references.pmid: 16207810
references.citation: Expert Rev Mol Med. 2005 Jul 18;7(13):1-19
references.pmid: 16026630
references.citation: Indian J Exp Biol. 1971 Jan;9(1):21-7
references.pmid: 4326108
references.citation: Biochem Biophys Res Commun. 1992 May 15;184(3):1261-5
references.pmid: 1317169
references.citation: Curr Opin Hematol. 1996 Jan;3(1):63-70
references.pmid: 9372053
references.citation: Adv Enzyme Regul. 2008;48:41-54
references.pmid: 18157946
references.citation: Exp Parasitol. 1994 Nov;79(3):424-35
references.pmid: 7957761
references.citation: Trans R Soc Trop Med Hyg. 1959 May;53(3):278-81
references.pmid: 13669033
references.citation: Trends Parasitol. 2012 Oct;28(10):417-26
references.pmid: 22906512
references.citation: Trans R Soc Trop Med Hyg. 1978;72(4):431-2
references.pmid: 212851
references.citation: J Leukoc Biol. 2006 Jun;79(6):1369-80
references.pmid: 16614259
references.citation: Arch Invest Med (Mex). 1974;5 SUPPL 2:423-6
references.pmid: 4376926
references.citation: Mol Biol Cell. 2007 Dec;18(12):5069-80
references.pmid: 17928408
references.citation: J Biol Chem. 1994 Jan 14;269(2):1518-22
references.pmid: 8288618
references.citation: Cell. 1995 Dec 15;83(6):1037-46
references.pmid: 8521504
references.citation: Mol Biol Cell. 2001 Nov;12(11):3550-62
references.pmid: 11694588
references.citation: Infect Immun. 1995 Nov;63(11):4358-67
references.pmid: 7591071
references.citation: Sci STKE. 2002 Apr 23;2002(129):pl6
references.pmid: 11972359
references.citation: Annu Rev Biophys Biomol Struct. 2002;31:151-75
references.pmid: 11988466