Endosomal recycling regulates Anthrax Toxin Receptor 1/Tumor Endothelial Marker 8-dependent cell spreading.
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.
Gu
Jingsheng
J
Department of Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322, USA.
Faundez
Victor
V
Werner
Erica
E
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
D008840:Microfilament Proteins; D009363:Neoplasm Proteins; D011956:Receptors, Cell Surface; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C473287:ANTXR1 protein, human
Mol Biol Cell. 1999 Dec;10(12):4163-76
10588650
J Biol Chem. 2006 Apr 28;281(17):12112-22
16507569
J Neurosci. 2002 Nov 15;22(22):9776-84
12427833
Front Biosci (Landmark Ed). 2009 Jan 01;14:4516-24
19273366
Nat Rev Mol Cell Biol. 2002 Aug;3(8):600-14
12154371
J Cell Sci. 2006 Sep 15;119(Pt 18):3723-31
16959902
Nat Rev Mol Cell Biol. 2004 Feb;5(2):121-32
15040445
Proc Natl Acad Sci U S A. 2004 May 4;101(18):7082-7
15105445
Traffic. 2004 Jan;5(1):20-36
14675422
Science. 1998 May 1;280(5364):734-7
9563949
J Biol Chem. 2003 Dec 5;278(49):49342-7
14507921
Proc Natl Acad Sci U S A. 2007 May 8;104(19):7939-44
17483462
Mol Biol Cell. 2008 Apr;19(4):1415-26
18256276
Proc Natl Acad Sci U S A. 1982 May;79(10):3162-6
6285339
Science. 2000 Aug 18;289(5482):1197-202
10947988
Mol Biol Cell. 2001 Jun;12(6):1843-57
11408590
PLoS One. 2007 Mar 28;2(3):e329
17389920
J Biol Chem. 2001 Nov 30;276(48):44712-20
11560936
Methods. 1998 Oct;16(2):150-9
9790861
J Cell Biol. 1999 Apr 5;145(1):123-39
10189373
J Cell Sci. 2007 Mar 1;120(Pt 5):802-14
17284518
J Biol Chem. 2003 Feb 14;278(7):5227-34
12468536
Cancer Res. 2001 Sep 15;61(18):6649-55
11559528
J Biol Chem. 2006 Aug 11;281(32):23227-36
16762926
J Biol Chem. 1984 Dec 10;259(23):14762-72
6094573
Curr Opin Cell Biol. 2006 Oct;18(5):549-57
16904305
Traffic. 2007 Dec;8(12):1854-1872
17944804
J Cell Biol. 2007 May 7;177(3):515-25
17485491
Annu Rev Biochem. 2007;76:243-65
17335404
Mol Biol Cell. 2004 Feb;15(2):575-87
14657250
J Biol Chem. 2008 Jan 4;283(1):529-540
17974567
Traffic. 2006 Jan;7(1):14-21
16445683
Proc Natl Acad Sci U S A. 1998 May 26;95(11):6187-92
9600939
Cancer Res. 2009 Aug 1;69(15):6021-6
19622764
J Cell Biol. 2003 Feb 3;160(3):321-8
12551953
Dev Cell. 2005 Aug;9(2):197-207
16054027
J Cell Biol. 2004 Aug 30;166(5):645-51
15337774
J Cell Biol. 2006 Jan 16;172(2):309-20
16401723
J Biol Chem. 2007 Aug 3;282(31):22513-24
17545147
Curr Opin Cell Biol. 1998 Aug;10(4):537-41
9719876
Cell. 1981 May;24(2):493-502
6263497
Mol Biol Cell. 2009 Jul;20(14):3261-72
19458190
J Cell Biol. 1996 Nov;135(4):913-24
8922376
Proc Natl Acad Sci U S A. 2003 Apr 29;100(9):5170-4
12700348
J Biol Chem. 2009 Jan 16;284(3):1790-802
19010779
J Cell Biol. 2002 Mar 4;156(5):843-54
11864992
Traffic. 2009 Aug;10(8):1115-27
19531065
Mol Biol Cell. 2009 Mar;20(5):1441-53
19144828
Cell Regul. 1991 Aug;2(8):599-612
1777504
J Cell Biol. 1998 Nov 16;143(4):957-71
9817754
Proc Natl Acad Sci U S A. 1998 Sep 15;95(19):11223-8
9736717
Mol Cell Biol. 2002 Oct;22(20):7279-90
12242303