title: Structure-function analysis of the WIP role in T cell receptor-stimulated NFAT activation: evidence that WIP-WASP dissociation is not required and that the WIP NH2 terminus is inhibitory.
abstract: WASP and its binding partner WIP play important roles in T cells both in actin polymerization and in interleukin-2 transcription. Aberrations thereof contribute to the pathology of Wiskott-Aldrich syndrome (WAS). To directly evaluate the cooperativity of WIP and WASP in interleukin-2 transcription, we investigated how the WIP-WASP complex regulates NF-AT-mediated gene transcription. We developed an improved model system for analysis, using WIP and WASP cotransfection into Jurkat cells, in which strong induction of NFAT reporter activation is observed with anti-T cell receptor (TCR) antibody without the phorbol 12-myristate 13-acetate usually used previously. Using this system, our findings contradict a prevailing conceptual model of TCR-induced WIP-WASP dissociation by showing in three ways that the WIP-WASP complex mediates TCR-induced NFAT activation without dissociation. First, phosphorylation of WIP Ser(488) does not cause dissociation of the WIP-WASP complex. Second, WIP-WASP complexes do not dissociate demonstrably after TCR stimulation. Third, a fusion protein of WIP to WASP efficiently mediates NFAT activation. Next, our studies clarify that WIP stabilization of WASP explains otherwise unexpected results in TCR-induced NFAT activation. Finally, we find that the NH(2) terminus of WIP is a highly inhibitory region for TCR-mediated transcriptional activation in which at least two elements contribute: the NH(2)-terminal polyproline and the NH(2)-terminal actin-binding WH2 domain. This suggests that WIP, like WASP, is subject to autoinhibition. Our data indicate that the WIP-WASP complex plays an important role in WASP stabilization and NFAT activation.
authors: Experimental Immunology Branch, NCI, National Institutes of Health, Bethesda, Maryland 20892-1360, USA.
authors: Xiaoyun
authors: X
authors: Dong
authors: Genaro
authors: G
authors: Patino-Lopez
authors: Fabio
authors: F
authors: Candotti
authors: Stephen
authors: S
authors: Shaw
mesh_terms: D000199:Actins / Q000737:chemistry; D017252:CD3 Complex / Q000096:biosynthesis; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism*; D019169:Jurkat Cells; D007700:Kinetics; D008954:Models, Biological; D050778:NFATC Transcription Factors / Q000737:chemistry / Q000502:physiology*; D017434:Protein Structure, Tertiary; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D013329:Structure-Activity Relationship; D013755:Tetradecanoylphorbol Acetate / Q000737:chemistry; D014158:Transcription, Genetic; D014923:Wiskott-Aldrich Syndrome / Q000378:metabolism; D051316:Wiskott-Aldrich Syndrome Protein / Q000378:metabolism*
chemical_list: D000199:Actins; D017252:CD3 Complex; D003598:Cytoskeletal Proteins; D007376:Interleukin-2; D047908:Intracellular Signaling Peptides and Proteins; D050778:NFATC Transcription Factors; D011948:Receptors, Antigen, T-Cell; C495767:WAS protein, human; C109903:WIPF1 protein, human; D051316:Wiskott-Aldrich Syndrome Protein; D013755:Tetradecanoylphorbol Acetate
references: J Biol Chem. 2001 Jun 15;276(24):21450-7
references: 11283014
references: Nat Cell Biol. 2000 Jul;2(7):441-8
references: 10878810
references: Eur J Immunol. 2000 Dec;30(12):3645-54
references: 11169407
references: J Immunol. 2001 Oct 15;167(8):4701-9
references: 11591801
references: Hum Gene Ther. 2002 Jun 10;13(9):1039-46
references: 12067437
references: Cell. 2002 Nov 15;111(4):565-76
references: 12437929
references: Mol Cell. 2002 Dec;10(6):1269-81
references: 12504004
references: Curr Opin Allergy Clin Immunol. 2003 Dec;3(6):427-36
references: 14612666
references: J Exp Med. 2004 Feb 2;199(3):357-68
references: 14757742
references: J Immunol. 2004 Aug 1;173(3):1658-62
references: 15265894
references: Cell. 1994 Aug 26;78(4):635-44
references: 8069912
references: Nat Genet. 1995 Apr;9(4):414-7
references: 7795648
references: Protein Sci. 1995 Dec;4(12):2455-68
references: 8580836
references: Cell. 1996 Mar 8;84(5):723-34
references: 8625410
references: J Clin Invest. 1996 Jun 1;97(11):2627-34
references: 8647957
references: Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14671-6
references: 9405671
references: Cell Mol Life Sci. 1998 Oct;54(10):1145-53
references: 9817992
references: J Immunol. 1999 Apr 15;162(8):5019-24
references: 10202051
references: J Immunol. 2005 Mar 1;174(5):2602-11
references: 15728466
references: FEBS J. 2005 Dec;272(23):6131-44
references: 16302976
references: J Allergy Clin Immunol. 2005 Dec;116(6):1364-71
references: 16337472
references: Biochem Biophys Res Commun. 2006 Feb 10;340(2):709-17
references: 16378591
references: Biochem Biophys Res Commun. 2006 Apr 7;342(2):529-36
references: 16488394
references: J Biol Chem. 2006 Apr 14;281(15):10589-97
references: 16403731
references: J Immunol. 2006 Nov 15;177(10):7451-61
references: 17082665
references: Curr Biol. 2006 Dec 5;16(23):2337-44
references: 17141616
references: Biochem Biophys Res Commun. 2007 Feb 23;353(4):875-81
references: 17207458
references: Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):926-31
references: 17213309
references: Int Immunol. 2007 Feb;19(2):185-92
references: 17205972
references: J Immunol. 2007 Mar 1;178(5):2987-95
references: 17312144
references: Biochem Biophys Res Commun. 2007 May 25;357(1):289-94
references: 17418095
references: J Immunol. 2000 Mar 15;164(6):2866-70
references: 10706671
references: Nature. 2000 Mar 9;404(6774):151-8
references: 10724160