title: Endosomal recycling regulates Anthrax Toxin Receptor 1/Tumor Endothelial Marker 8-dependent cell spreading.
abstract: Mechanisms for receptor-mediated anthrax toxin internalization and delivery to the cytosol are well understood. However, far less is known about the fate followed by anthrax toxin receptors prior and after cell exposure to the toxin. We report that Anthrax Toxin Receptor 1/Tumor Endothelial Marker 8 (TEM8) localized at steady state in Rab11a-positive and transferrin receptor-containing recycling endosomes. TEM8 followed a slow constitutive recycling route of approximately 30min as determined by pulsed surface biotinylation and chase experiments. A Rab11a dominant negative mutant and Myosin Vb tail expression impaired TEM8 recycling by sequestering TEM8 in intracellular compartments. Sequestration of TEM8 in intracellular compartments with monensin coincided with increased TEM8 association with a multi-protein complex isolated with antibodies against transferrin receptor. Addition of the cell-binding component of anthrax toxin, Protective Antigen, reduced TEM8 half-life from 7 to 3 hours, without preventing receptor recycling. Pharmacological and molecular perturbation of recycling endosome function using monensin, dominant negative Rab11a, or myosin Vb tail, reduced PA binding efficiency and TEM8-dependent cell spreading on PA-coated surfaces without affecting toxin delivery to the cytosol. These results indicate that the intracellular fate of TEM8 differentially affect its cell adhesion and cell intoxication functions.
authors.affiliation: Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
authors.forename: Jingsheng
authors.initials: J
authors.lastname: Gu
authors.forename: Victor
authors.initials: V
authors.lastname: Faundez
authors.forename: Erica
authors.initials: E
authors.lastname: Werner
mesh_terms: D002478:Cells, Cultured; D011992:Endosomes / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D008840:Microfilament Proteins; D009363:Neoplasm Proteins / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000099193:rab11 GTP-Binding Proteins
chemical_list: D008840:Microfilament Proteins; D009363:Neoplasm Proteins; D011956:Receptors, Cell Surface; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C473287:ANTXR1 protein, human
references.citation: Mol Biol Cell. 1999 Dec;10(12):4163-76
references.pmid: 10588650
references.citation: J Biol Chem. 2006 Apr 28;281(17):12112-22
references.pmid: 16507569
references.citation: J Neurosci. 2002 Nov 15;22(22):9776-84
references.pmid: 12427833
references.citation: Front Biosci (Landmark Ed). 2009 Jan 01;14:4516-24
references.pmid: 19273366
references.citation: Nat Rev Mol Cell Biol. 2002 Aug;3(8):600-14
references.pmid: 12154371
references.citation: J Cell Sci. 2006 Sep 15;119(Pt 18):3723-31
references.pmid: 16959902
references.citation: Nat Rev Mol Cell Biol. 2004 Feb;5(2):121-32
references.pmid: 15040445
references.citation: Proc Natl Acad Sci U S A. 2004 May 4;101(18):7082-7
references.pmid: 15105445
references.citation: Traffic. 2004 Jan;5(1):20-36
references.pmid: 14675422
references.citation: Science. 1998 May 1;280(5364):734-7
references.pmid: 9563949
references.citation: J Biol Chem. 2003 Dec 5;278(49):49342-7
references.pmid: 14507921
references.citation: Proc Natl Acad Sci U S A. 2007 May 8;104(19):7939-44
references.pmid: 17483462
references.citation: Mol Biol Cell. 2008 Apr;19(4):1415-26
references.pmid: 18256276
references.citation: Proc Natl Acad Sci U S A. 1982 May;79(10):3162-6
references.pmid: 6285339
references.citation: Science. 2000 Aug 18;289(5482):1197-202
references.pmid: 10947988
references.citation: Mol Biol Cell. 2001 Jun;12(6):1843-57
references.pmid: 11408590
references.citation: PLoS One. 2007 Mar 28;2(3):e329
references.pmid: 17389920
references.citation: J Biol Chem. 2001 Nov 30;276(48):44712-20
references.pmid: 11560936
references.citation: Methods. 1998 Oct;16(2):150-9
references.pmid: 9790861
references.citation: J Cell Biol. 1999 Apr 5;145(1):123-39
references.pmid: 10189373
references.citation: J Cell Sci. 2007 Mar 1;120(Pt 5):802-14
references.pmid: 17284518
references.citation: J Biol Chem. 2003 Feb 14;278(7):5227-34
references.pmid: 12468536
references.citation: Cancer Res. 2001 Sep 15;61(18):6649-55
references.pmid: 11559528
references.citation: J Biol Chem. 2006 Aug 11;281(32):23227-36
references.pmid: 16762926
references.citation: J Biol Chem. 1984 Dec 10;259(23):14762-72
references.pmid: 6094573
references.citation: Curr Opin Cell Biol. 2006 Oct;18(5):549-57
references.pmid: 16904305
references.citation: Traffic. 2007 Dec;8(12):1854-1872
references.pmid: 17944804
references.citation: J Cell Biol. 2007 May 7;177(3):515-25
references.pmid: 17485491
references.citation: Annu Rev Biochem. 2007;76:243-65
references.pmid: 17335404
references.citation: Mol Biol Cell. 2004 Feb;15(2):575-87
references.pmid: 14657250
references.citation: J Biol Chem. 2008 Jan 4;283(1):529-540
references.pmid: 17974567
references.citation: Traffic. 2006 Jan;7(1):14-21
references.pmid: 16445683
references.citation: Proc Natl Acad Sci U S A. 1998 May 26;95(11):6187-92
references.pmid: 9600939
references.citation: Cancer Res. 2009 Aug 1;69(15):6021-6
references.pmid: 19622764
references.citation: J Cell Biol. 2003 Feb 3;160(3):321-8
references.pmid: 12551953
references.citation: Dev Cell. 2005 Aug;9(2):197-207
references.pmid: 16054027
references.citation: J Cell Biol. 2004 Aug 30;166(5):645-51
references.pmid: 15337774
references.citation: J Cell Biol. 2006 Jan 16;172(2):309-20
references.pmid: 16401723
references.citation: J Biol Chem. 2007 Aug 3;282(31):22513-24
references.pmid: 17545147
references.citation: Curr Opin Cell Biol. 1998 Aug;10(4):537-41
references.pmid: 9719876
references.citation: Cell. 1981 May;24(2):493-502
references.pmid: 6263497
references.citation: Mol Biol Cell. 2009 Jul;20(14):3261-72
references.pmid: 19458190
references.citation: J Cell Biol. 1996 Nov;135(4):913-24
references.pmid: 8922376
references.citation: Proc Natl Acad Sci U S A. 2003 Apr 29;100(9):5170-4
references.pmid: 12700348
references.citation: J Biol Chem. 2009 Jan 16;284(3):1790-802
references.pmid: 19010779
references.citation: J Cell Biol. 2002 Mar 4;156(5):843-54
references.pmid: 11864992
references.citation: Traffic. 2009 Aug;10(8):1115-27
references.pmid: 19531065
references.citation: Mol Biol Cell. 2009 Mar;20(5):1441-53
references.pmid: 19144828
references.citation: Cell Regul. 1991 Aug;2(8):599-612
references.pmid: 1777504
references.citation: J Cell Biol. 1998 Nov 16;143(4):957-71
references.pmid: 9817754
references.citation: Proc Natl Acad Sci U S A. 1998 Sep 15;95(19):11223-8
references.pmid: 9736717
references.citation: Mol Cell Biol. 2002 Oct;22(20):7279-90
references.pmid: 12242303