Proximal human FOXP3 promoter transactivated by NF-kappaB and negatively controlled by feedback loop and SP3.
Forkhead box protein 3 (Foxp3) is indispensable for the development of CD4(+)CD25(+) regulatory T cells (Tregs). Here we analyzed three prominent evolutionary conserved regions (ECRs) upstream of the transcription start site of the human FOXP3 gene. We show that ECR2 and ECR3 fragments derived from positions -1.3 to -2.0 kb and -5.0 to -6.0 kb, respectively, display basal transcriptional activity. Reporter constructs derived from ECR1, located between -0.6 and +0.23 kb and thus the most proximal ECR in respect of transcription initiation, remained almost inactive. However, ECR1 was transactivated by the NF-kappaB subunit p65 in HEK 293 cells. In Jurkat and primary T cells, in addition to p65, a second stimulus delivered by either T-cell receptor stimulation or addition of PMA was needed. Co-expression of I kappaB alpha inhibited p65-mediated FOXP3 proximal promoter transactivation, and the NF-kappaB inhibitor curcumin reduced Foxp3 neoexpression in IL-2/CD3/CD28/TGF-beta stimulated PBMCs. Moreover, proximal FOXP3 promoter transactivation was inhibited by Foxp3 and the SP transcription factor family member SP3. Thus, the human proximal FOXP3 promoter is controlled by activation through the TCR involving PKC and the NF-kappaB subunit p65 and by inhibition through a negative feedback loop and SP3.
Eckerstorfer
Paul
P
Molecular Immunology Unit, Institute for Hygiene and Applied Immunology, Center for Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Lazarettgasse 19, A-1090 Vienna, Austria.
Novy
Michael
M
Burgstaller-Muehlbacher
Sebastian
S
Paster
Wolfgang
W
Schiller
Herbert B
HB
Mayer
Herbert
H
Stockinger
Hannes
H
D002460:Cell Line; D003474:Curcumin / Q000494:pharmacology; D025461:Feedback, Physiological; D051858:Forkhead Transcription Factors / Q000235:genetics*; D006801:Humans; D011401:Promoter Regions, Genetic; D051705:Sp3 Transcription Factor / Q000502:physiology*; D050378:T-Lymphocytes, Regulatory / Q000378:metabolism; D051996:Transcription Factor RelA / Q000502:physiology*; D015533:Transcriptional Activation
C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; C497121:SP3 protein, human; D051996:Transcription Factor RelA; D051705:Sp3 Transcription Factor; D003474:Curcumin
Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W249-52
15215389
Eur J Immunol. 2007 Jan;37(1):129-38
17154262
J Biol Chem. 2005 Jan 14;280(2):1217-23
15536066
J Immunol. 2003 Oct 1;171(7):3435-41
14500638
J Clin Invest. 2000 Dec;106(12):R75-81
11120765
Genome Res. 2002 Jun;12(6):996-1006
12045153
J Exp Med. 2005 Jan 17;201(2):181-7
15657288
Proc Natl Acad Sci U S A. 2004 Mar 30;101(13):4566-71
15070758
Proc Natl Acad Sci U S A. 2005 Apr 5;102(14):5138-43
15790681
Annu Rev Immunol. 2006;24:209-26
16551248
Immunity. 2009 Dec 18;31(6):932-40
20064450
Eur J Immunol. 2009 Sep;39(9):2571-83
19701891
Mol Biol Cell. 2005 May;16(5):2181-90
15716352
Tohoku J Exp Med. 2006 Sep;210(1):1-9
16960339
J Exp Med. 2009 Dec 21;206(13):3001-14
19995950
Hum Immunol. 2005 Jan;66(1):13-20
15620457
Eur J Immunol. 2009 Jan;39(1):78-84
19130560
Nature. 2007 Feb 22;445(7130):936-40
17237761
J Exp Med. 2007 Jul 9;204(7):1543-51
17591856
Nat Genet. 2001 Jan;27(1):20-1
11137993
EMBO J. 1994 Aug 15;13(16):3843-51
8070411
Blood. 2006 Sep 1;108(5):1571-9
16645171
Mol Cell Biol. 1999 Mar;19(3):2098-108
10022897
PLoS Biol. 2009 Mar 3;7(3):e51
19260764
Nat Rev Mol Cell Biol. 2001 Nov;2(11):827-37
11715049
J Immunol. 2008 Nov 15;181(10):6898-905
18981109
J Surg Res. 2004 Oct;121(2):171-7
15501456
Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W280-6
15215395
J Immunol. 2007 Jan 1;178(1):280-90
17182565
PLoS Biol. 2007 Feb;5(2):e38
17298177
Nat Immunol. 2008 Feb;9(2):194-202
18157133
Nat Rev Mol Cell Biol. 2002 Sep;3(9):651-62
12209125
Biochem Biophys Res Commun. 2007 Oct 12;362(1):44-50
17706604
J Clin Invest. 2005 Nov;115(11):3276-84
16211090
Nat Genet. 2001 Jan;27(1):68-73
11138001
Mol Immunol. 2008 Dec;46(2):213-24
18842300
J Biol Chem. 2001 Oct 5;276(40):37672-9
11483607
Immunity. 2003 Sep;19(3):377-89
14499113
J Med Genet. 2002 Aug;39(8):537-45
12161590
J Immunol. 2009 Jun 1;182(11):6736-43
19454668
Nature. 2007 Feb 22;445(7130):931-5
17237765
Nat Immunol. 2007 Mar;8(3):277-84
17220892
J Immunol. 2017 Feb 1;198(3):981-985
28115586
Nature. 2007 Apr 5;446(7136):685-9
17377532
Blood. 2007 Oct 15;110(8):2983-90
17644734
J Immunol. 2009 Jan 15;182(2):1041-9
19124747
Genes Dev. 2004 Sep 15;18(18):2195-224
15371334
Nucleic Acids Res. 1992 Jun 11;20(11):2902
1614888
Eur J Immunol. 1997 Jul;27(7):1601-9
9247567
Immunogenetics. 2003 Jun;55(3):149-156
12750858
J Exp Med. 2003 Dec 15;198(12):1875-86
14676299
Nat Rev Mol Cell Biol. 2002 Jan;3(1):11-20
11823794
J Immunol. 2006 Mar 15;176(6):3593-602
16517728
Immunity. 2009 Dec 18;31(6):921-31
20064449
Nat Med. 2004 Sep;10(9):900-1
15340410