title: Pseudomonas aeruginosa exotoxin Y-mediated tau hyperphosphorylation impairs microtubule assembly in pulmonary microvascular endothelial cells.
abstract: Pseudomonas aeruginosa uses a type III secretion system to introduce the adenylyl and guanylyl cyclase exotoxin Y (ExoY) into the cytoplasm of endothelial cells. ExoY induces Tau hyperphosphorylation and insolubility, microtubule breakdown, barrier disruption and edema, although the mechanism(s) responsible for microtubule breakdown remain poorly understood. Here we investigated both microtubule behavior and centrosome activity to test the hypothesis that ExoY disrupts microtubule dynamics. Fluorescence microscopy determined that infected pulmonary microvascular endothelial cells contained fewer microtubules than control cells, and further studies demonstrated that the microtubule-associated protein Tau was hyperphosphorylated following infection and dissociated from microtubules. Disassembly/reassembly studies determined that microtubule assembly was disrupted in infected cells, with no detectable effects on either microtubule disassembly or microtubule nucleation by centrosomes. This effect of ExoY on microtubules was abolished when the cAMP-dependent kinase phosphorylation site (Ser-214) on Tau was mutated to a non-phosphorylatable form. These studies identify Tau in microvascular endothelial cells as the target of ExoY in control of microtubule architecture following pulmonary infection by Pseudomonas aeruginosa and demonstrate that phosphorylation of tau following infection decreases microtubule assembly.
authors.affiliation: Department of Cell Biology and Neuroscience, University of South Alabama, Mobile, Alabama, United States of America ; Center for Lung Biology, University of South Alabama, Mobile, Alabama, United States of America.
authors.forename: Ron
authors.initials: R
authors.lastname: Balczon
authors.forename: Nutan
authors.initials: N
authors.lastname: Prasain
authors.forename: Cristhiaan
authors.initials: C
authors.lastname: Ochoa
authors.forename: Jason
authors.initials: J
authors.lastname: Prater
authors.forename: Bing
authors.initials: B
authors.lastname: Zhu
authors.forename: Mikhail
authors.initials: M
authors.lastname: Alexeyev
authors.forename: Sarah
authors.initials: S
authors.lastname: Sayner
authors.forename: Dara W
authors.initials: DW
authors.lastname: Frank
authors.forename: Troy
authors.initials: T
authors.lastname: Stevens
mesh_terms: D000818:Animals; D018385:Centrosome / Q000187:drug effects / Q000378:metabolism; D042783:Endothelial Cells / Q000166:cytology* / Q000187:drug effects; D005098:Exotoxins / Q000633:toxicity*; D008168:Lung / Q000098:blood supply*; D008870:Microtubules / Q000187:drug effects* / Q000378:metabolism; D055806:Microvessels / Q000166:cytology*; D010766:Phosphorylation / Q000187:drug effects; D011550:Pseudomonas aeruginosa / Q000378:metabolism*; D051381:Rats; D012694:Serine / Q000378:metabolism; D016875:tau Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D005098:Exotoxins; D016875:tau Proteins; D012694:Serine
references.citation: Am J Physiol Lung Cell Mol Physiol. 2011 Aug;301(2):L157-60
references.pmid: 21685243
references.citation: Circ Res. 2004 Jul 23;95(2):196-203
references.pmid: 15192021
references.citation: J Cell Sci. 2001 Feb;114(Pt 4):797-809
references.pmid: 11171385
references.citation: Curr Opin Genet Dev. 2001 Jun;11(3):343-51
references.pmid: 11377973
references.citation: J Gen Physiol. 2000 Aug;116(2):147-61
references.pmid: 10919863
references.citation: Am J Physiol. 1999 Jul;277(1):L119-26
references.pmid: 10409238
references.citation: Curr Opin Cell Biol. 1995 Feb;7(1):72-81
references.pmid: 7755992
references.citation: Mol Aspects Med. 2009 Dec;30(6):423-30
references.pmid: 19560485
references.citation: Proc Natl Acad Sci U S A. 1998 Nov 10;95(23):13899-904
references.pmid: 9811898
references.citation: J Cell Sci. 1993 May;105 ( Pt 1):51-60
references.pmid: 7689576
references.citation: EMBO J. 2006 May 17;25(10):2051-61
references.pmid: 16642035
references.citation: Proc Natl Acad Sci U S A. 1982 May;79(10):3162-6
references.pmid: 6285339
references.citation: Circ Res. 2006 Mar 17;98(5):675-81
references.pmid: 16469954
references.citation: J Clin Invest. 1999 Sep;104(6):743-50
references.pmid: 10491409
references.citation: Curr Opin Infect Dis. 2003 Apr;16(2):135-43
references.pmid: 12734446
references.citation: Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2696-700
references.pmid: 7708708
references.citation: Am J Physiol Lung Cell Mol Physiol. 2012 Feb 1;302(3):L275-86
references.pmid: 21964402
references.citation: Nucleic Acids Res. 2009 Jan;37(Database issue):D175-80
references.pmid: 18971254
references.citation: J Cell Physiol. 1989 Apr;139(1):157-66
references.pmid: 2540209
references.citation: EMBO J. 2005 Sep 21;24(18):3190-201
references.pmid: 16138079
references.citation: Am J Physiol Cell Physiol. 2012 Mar 15;302(6):C839-52
references.pmid: 22116306
references.citation: Am J Physiol. 1998 Aug;275(2):L203-22
references.pmid: 9700080
references.citation: Am J Physiol Lung Cell Mol Physiol. 2009 Jul;297(1):L73-83
references.pmid: 19395666
references.citation: J Gen Physiol. 2001 Jul;118(1):63-78
references.pmid: 11429444
references.citation: Science. 2001 Sep 21;293(5538):2204-5
references.pmid: 11569497
references.citation: Am J Physiol. 1997 Jul;273(1 Pt 1):L22-30
references.pmid: 9252536
references.citation: N Engl J Med. 2010 Jan 28;362(4):329-44
references.pmid: 20107219
references.citation: Nature. 2003 Dec 4;426(6966):570-4
references.pmid: 14654843
references.citation: Thorax. 1998 Mar;53(3):213-9
references.pmid: 9659359
references.citation: Eur J Biochem. 1998 Mar 15;252(3):542-52
references.pmid: 9546672
references.citation: J Cell Sci. 2008 Jan 1;121(Pt 1):110-9
references.pmid: 18073242
references.citation: FASEB J. 2004 Dec;18(15):1879-90
references.pmid: 15576491
references.citation: Cell Biochem Funct. 1995 Dec;13(4):239-50
references.pmid: 10232926
references.citation: Science. 2008 Nov 28;322(5906):1353-6
references.pmid: 18927356
references.citation: Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10369-73
references.pmid: 7479786
references.citation: Am J Physiol Lung Cell Mol Physiol. 2010 Oct;299(4):L493-501
references.pmid: 20639351
references.citation: J Cell Physiol. 2005 Sep;204(3):934-47
references.pmid: 15828024
references.citation: J Cell Sci. 1992 Dec;103 ( Pt 4):965-76
references.pmid: 1487507
references.citation: Biochemistry. 2010 Jul 6;49(26):5494-503
references.pmid: 20521845
references.citation: Am J Physiol Lung Cell Mol Physiol. 2001 Sep;281(3):L565-74
references.pmid: 11504682
references.citation: Am J Physiol Lung Cell Mol Physiol. 2011 Jul;301(1):L117-24
references.pmid: 21478251
references.citation: J Cell Biol. 1988 Apr;106(4):1213-20
references.pmid: 3283150
references.citation: Trends Biochem Sci. 2009 Jul;34(7):332-42
references.pmid: 19559622
references.citation: J Biol Chem. 2012 Jul 20;287(30):25407-18
references.pmid: 22637478
references.citation: N Engl J Med. 2010 Sep 30;363(14):1293-6
references.pmid: 20879875
references.citation: Thromb Haemost. 2001 Jul;86(1):308-15
references.pmid: 11487019
references.citation: J Appl Physiol (1985). 2001 Oct;91(4):1487-500
references.pmid: 11568129
references.citation: J Neurosci. 2007 Aug 22;27(34):9155-68
references.pmid: 17715352
references.citation: J Cell Biol. 2002 Jun 24;157(7):1267-78
references.pmid: 12082084
references.citation: Am J Forensic Med Pathol. 2010 Jun;31(2):130-2
references.pmid: 20032774
references.citation: Can J Neurol Sci. 2011 Jul;38(4):547-56
references.pmid: 21672694
references.citation: Am J Physiol Lung Cell Mol Physiol. 2011 Jan;300(1):L132-8
references.pmid: 20971804
references.citation: J Biol Chem. 1979 Dec 25;254(24):12670-8
references.pmid: 227904
references.citation: Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):13049-54
references.pmid: 11606735
references.citation: Proc Natl Acad Sci U S A. 2003 Aug 5;100(16):9548-53
references.pmid: 12886013
references.citation: J Biol Chem. 2007 Nov 30;282(48):34801-8
references.pmid: 17921144
references.citation: Am J Physiol. 1989 Sep;257(3 Pt 1):C562-7
references.pmid: 2782396