title: Pasteurella multocida toxin as a transporter of non-cell-permeating proteins.
abstract: The protein toxin Pasteurella multocida toxin (PMT) is the causative agent of atrophic rhinitis in pigs, leading to atrophy of the nasal turbinate bones by affecting osteoblasts and osteoclasts. The mechanism of PMT-induced intoxication is a deamidation of α-subunits of heterotrimeric G proteins, including Gαq, Gα13, and Gαi, thereby causing persistent activation of the G proteins. Here we utilized PMT as a transporter of the non-cell-permeating A domain of diphtheria toxin (DTa). Fusion proteins of PMT and DTa ADP-ribosylated elongation factor 2, the natural target of diphtheria toxin, leading to cell toxicity. PMT-DTa effects were competed by PMT, indicating binding to the same cell surface receptor. Fluorescently labeled PMT-DTa and PMT colocalized with specific markers of early and late endosomes. Bafilomycin A, which inhibits vacuolar H(+)-ATPase, blocked PMT-DTa-induced intoxication of HEK-293 cells. By constructing various PMT-DTa chimeras, we identified a minimal region of PMT necessary for uptake of DTa. The data suggest that PMT is able to transport cargo proteins into eukaryotic cells by utilizing the PMT-specific uptake route.
authors: Institut für Experimentelle und Klinische Pharmakologie und Toxikologie, Albert-Ludwigs-Universität Freiburg, Freiburg, Germany.
authors: Stefan
authors: S
authors: Bergmann
authors: Doris
authors: D
authors: Jehle
authors: Carsten
authors: C
authors: Schwan
authors: Joachim H C
authors: JH
authors: Orth
authors: Klaus
authors: K
authors: Aktories
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D018938:Caco-2 Cells; D002463:Cell Membrane Permeability; D002470:Cell Survival; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism; D004167:Diphtheria Toxin / Q000235:genetics / Q000378:metabolism*; D011992:Endosomes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D018942:Macrolides / Q000494:pharmacology; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D001427:Bacterial Toxins; D015415:Biomarkers; D004167:Diphtheria Toxin; D018942:Macrolides; C062258:Pasteurella multocida toxin; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C029795:diphtheria toxin fragment A; C057620:bafilomycin A
references: J Biol Chem. 1991 Sep 15;266(26):17707-12
references: 1832676
references: Cell Signal. 2009 Apr;21(4):551-8
references: 19135527
references: Open Biochem J. 2007;1:7-11
references: 18949067
references: Cell Microbiol. 2007 Oct;9(10):2485-96
references: 17581254
references: Restor Neurol Neurosci. 1995 Jan 1;8(1):13-4
references: 21551798
references: Biochemistry. 2003 May 6;42(17):4971-7
references: 12718539
references: Naunyn Schmiedebergs Arch Pharmacol. 2002 Dec;366(6):501-12
references: 12444490
references: Bacteriol Rev. 1975 Mar;39(1):54-85
references: 164179
references: J Biol Chem. 1971 Mar 10;246(5):1504-10
references: 5545093
references: FEMS Microbiol Lett. 2006 Dec;265(1):1-10
references: 17107417
references: FASEB J. 2013 Feb;27(2):832-42
references: 23150526
references: Proc Natl Acad Sci U S A. 2007 Mar 20;104(12):5139-44
references: 17360394
references: Cell Microbiol. 2010 Aug;12(8):1174-85
references: 20331638
references: Mol Microbiol. 2004 Oct;54(1):239-50
references: 15458419
references: FEBS J. 2011 Dec;278(23):4633-48
references: 21951695
references: Int J Med Microbiol. 2001 Sep;291(4):261-8
references: 11680786
references: Curr Top Microbiol Immunol. 2012;361:73-92
references: 22371145
references: Science. 1969 Jun 6;164(3884):1179-81
references: 4305968
references: Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12424-9
references: 10535938
references: Curr Top Microbiol Immunol. 2012;361:113-29
references: 22411430
references: J Biol Chem. 2008 Aug 22;283(34):23288-94
references: 18583341
references: J Biol Chem. 1997 Jan 10;272(2):1268-75
references: 8995431
references: Curr Top Microbiol Immunol. 2012;361:1-22
references: 22643916
references: J Am Acad Dermatol. 1995 Dec;33(6):1019-29
references: 7490347
references: Zentralbl Veterinarmed B. 1991 Jul;38(5):345-52
references: 1927149
references: J Biol Chem. 2010 Aug 13;285(33):25467-75
references: 20534589
references: J Biol Chem. 2001 Feb 9;276(6):3840-5
references: 11062245
references: Curr Top Microbiol Immunol. 2012;361:23-38
references: 22402726
references: J Biol Chem. 2005 Nov 4;280(44):36701-7
references: 16141214
references: Nature. 2011 Jul 19;477(7366):549-55
references: 21772288
references: Nat Chem Biol. 2007 Nov;3(11):707-8
references: 17891153
references: Annu Rev Microbiol. 2008;62:271-88
references: 18785839
references: Nature. 1992 May 21;357(6375):216-22
references: 1589020
references: Methods Enzymol. 1995;256:297-306
references: 7476444
references: J Am Chem Soc. 2009 Aug 12;131(31):10800-1
references: 19610623
references: Cell. 1997 Apr 18;89(2):251-61
references: 9108480
references: J Biol Chem. 1971 Mar 10;246(5):1496-503
references: 5545092
references: Infect Immun. 1998 Dec;66(12):5636-42
references: 9826336
references: Infect Immun. 2001 Jun;69(6):3628-34
references: 11349023
references: Curr Opin Cell Biol. 2008 Aug;20(4):415-26
references: 18511251
references: Infect Immun. 2009 May;77(5):1835-41
references: 19237521
references: Proc Natl Acad Sci U S A. 1990 Jan;87(1):123-7
references: 2153282
references: Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):7179-84
references: 19369209
references: J Biol Chem. 1968 Jun 25;243(12):3553-5
references: 4297784
references: Proc Natl Acad Sci U S A. 1996 Aug 6;93(16):8437-42
references: 8710889
references: Proc Natl Acad Sci U S A. 2010 Mar 23;107(12):5581-6
references: 20212166
references: Curr Opin Cell Biol. 2000 Aug;12(4):407-13
references: 10873820
references: Naunyn Schmiedebergs Arch Pharmacol. 2011 Mar;383(3):263-73
references: 21136248
references: Toxins (Basel). 2011 Mar;3(3):218-41
references: 22053287
references: J Exp Med. 1969 Jan 1;129(1):1-21
references: 4304436