Salmonella type III secretion effector SlrP is an E3 ubiquitin ligase for mammalian thioredoxin.
Salmonella enterica encodes two virulence-related type III secretion systems in Salmonella pathogenicity islands 1 and 2, respectively. These systems mediate the translocation of protein effectors into the eukaryotic host cell, where they alter cell signaling and manipulate host cell functions. However, the precise role of most effectors remains unknown. Using a genetic screen, we identified the small, reduction/oxidation-regulatory protein thioredoxin as a mammalian binding partner of the Salmonella effector SlrP. The interaction was confirmed by affinity chromatography and coimmunoprecipitation. In vitro, SlrP was able to mediate ubiquitination of ubiquitin and thioredoxin. A Cys residue conserved in other effectors of the same family that also possess E3 ubiquitin ligase activity was essential for this catalytic function. Stable expression of SlrP in HeLa cells resulted in a significant decrease of thioredoxin activity and in an increase of cell death. The physiological significance of these results was strengthened by the finding that Salmonella was able to trigger cell death and inhibit thioredoxin activity in HeLa cells several hours post-infection. This study assigns a functional role to the Salmonella effector SlrP as a binding partner and an E3 ubiquitin ligase for mammalian thioredoxin.
Bernal-Bayard
Joaquín
J
Departamento de Genética, Facultad de Biología, Universidad de Sevilla, 41080 Sevilla, Spain.
Ramos-Morales
Francisco
F
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D016923:Cell Death; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D047468:Immunoprecipitation; D011485:Protein Binding / Q000187:drug effects; D012475:Salmonella; D012480:Salmonella Infections / Q000378:metabolism / Q000473:pathology; D013879:Thioredoxins / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*
D001426:Bacterial Proteins; C401846:SlrP protein, Salmonella typhimurium; D013879:Thioredoxins; D006861:Hydrogen Peroxide; D044767:Ubiquitin-Protein Ligases
Proc Natl Acad Sci U S A. 2001 Dec 18;98(26):15264-9
11742086
Trends Microbiol. 2007 Nov;15(11):516-24
17983751
Methods Enzymol. 1999;300:226-39
9919525
Curr Top Microbiol Immunol. 1996;209:43-60
8742245
J Exp Med. 1994 Jul 1;180(1):15-23
8006579
Semin Cancer Biol. 2006 Dec;16(6):420-6
17092741
Free Radic Biol Med. 2001 Dec 1;31(11):1287-312
11728801
Cell. 1993 Nov 19;75(4):805-16
8242751
Infect Immun. 2005 Oct;73(10):6260-71
16177297
Cell Microbiol. 2002 Nov;4(11):771-81
12427099
Proc Natl Acad Sci U S A. 2009 Mar 24;106(12):4864-9
19273841
J Mol Biol. 2005 May 13;348(4):817-30
15843015
Science. 1993 May 28;260(5112):1338-43
8493579
Cell Microbiol. 2007 Nov;9(11):2562-70
17714514
Blood. 2005 Feb 15;105(4):1598-605
15494431
J Biol Chem. 2005 Mar 11;280(10):9058-64
15642738
Nat Struct Mol Biol. 2008 Dec;15(12):1293-301
18997778
Cell Microbiol. 2008 Feb;10(2):285-92
18034865
J Immunol. 2002 Sep 15;169(6):2846-50
12218096
EMBO J. 1998 May 1;17(9):2596-606
9564042
Nat Rev Microbiol. 2006 Nov;4(11):811-25
17041629
Antioxid Redox Signal. 2005 Sep-Oct;7(9-10):1189-94
16115022
J Bioenerg Biomembr. 2003 Apr;35(2):169-79
12887015
Cell Host Microbe. 2007 Mar 15;1(1):77-83
18005683
Annu Rev Biochem. 1985;54:237-71
3896121
Mol Cell Biol. 2003 Feb;23(3):916-22
12529397
Cell Mol Life Sci. 2008 Aug;65(15):2397-406
18438605
Microbiol Mol Biol Rev. 2004 Dec;68(4):771-95
15590783
Infect Immun. 1999 Dec;67(12):6385-93
10569754
Dev Biol. 1996 Aug 25;178(1):179-85
8812119
J Biol Chem. 1999 Jul 30;274(31):21645-50
10419473
Curr Opin Immunol. 2001 Aug;13(4):410-6
11498296
J Bacteriol. 2003 Sep;185(17):5096-108
12923082
Annu Rev Biochem. 2001;70:503-33
11395416
J Exp Med. 1999 Jun 7;189(11):1783-9
10359582
Annu Rev Biophys Biomol Struct. 2001;30:421-55
11441809
J Immunol. 2000 Jun 15;164(12):6287-95
10843682
Cell Microbiol. 2007 Sep;9(9):2103-11
17593246
Nat Cell Biol. 2004 Jul;6(7):634-41
15208638
J Mol Biol. 1983 Jun 5;166(4):557-80
6345791
J Biol Chem. 1964 Oct;239:3436-44
14245400
Nat Cell Biol. 2002 Dec;4(12):E295-8
12461537
J Clin Invest. 1998 Nov 15;102(10):1815-23
9819367
Mol Microbiol. 1997 May;24(4):697-709
9194698
Int J Med Microbiol. 2007 Oct;297(6):401-15
17482513
J Biol Chem. 1999 Sep 24;274(39):27891-7
10488136
Nat Struct Mol Biol. 2008 Dec;15(12):1302-8
18997779
J Cell Biochem. 2008 Aug 1;104(5):1879-89
18384140
Cell Microbiol. 2006 May;8(5):837-46
16611232
Cell Mol Life Sci. 2004 Nov;61(22):2812-26
15558211
Trends Biochem Sci. 2000 Nov;25(11):544-8
11084366
Biochemistry. 1982 Dec 21;21(26):6628-33
7159551
Free Radic Biol Med. 1995 Jul;19(1):53-65
7635359
Biochemistry. 1997 Nov 18;36(46):13979-88
9369469
Eur J Biochem. 2000 Oct;267(20):6102-9
11012661
PLoS Pathog. 2007 Jan;3(1):e3
17257058
Nat Genet. 1997 Jul;16(3):277-82
9207794
Proc Natl Acad Sci U S A. 2000 Jun 20;97(13):7539-44
10861017
J Bacteriol. 1999 Jun;181(11):3433-7
10348855
Microbes Infect. 2001 Nov-Dec;3(14-15):1321-6
11755421
Structure. 1996 Jun 15;4(6):735-51
8805557
Microbiology (Reading). 2008 Jun;154(Pt 6):1570-1583
18524912