IL-10 inhibits transcription elongation of the human TNF gene in primary macrophages.
IL-10 plays a central nonredundant role in limiting inflammation in vivo. However, the mechanisms involved remain to be resolved. Using primary human macrophages, we found that IL-10 inhibits selected inflammatory genes, primarily at a level of transcription. At the TNF gene, this occurs not through an inhibition of RNA polymerase II (Pol II) recruitment and transcription initiation but through a mechanism targeting the stimulation of transcription elongation by cyclin-dependent kinase (CDK) 9. We demonstrated an unanticipated requirement for a region downstream of the TNF 3' untranslated region (UTR) that contains the nuclear factor κB (NF-κB) binding motif (κB4) both for induction of transcription by lipopolysaccharide (LPS) and its inhibition by IL-10. IL-10 not only inhibits the recruitment of RelA to regions containing κB sites at the TNF gene but also to those found at other LPS-induced genes. We show that although IL-10 elicits a general block in RelA recruitment to its genomic targets, the gene-specific nature of IL-10's actions are defined through the differential recruitment of CDK9 and the control of transcription elongation. At TNF, but not NFKBIA, the consequence of RelA recruitment inhibition is a loss of CDK9 recruitment, preventing the stimulation of transcription elongation.
Smallie
Tim
T
Kennedy Institute of Rheumatology Division, Imperial College London, London W6 8LH, UK.
Ricchetti
Giuseppe
G
Horwood
Nicole J
NJ
Feldmann
Marc
M
Clark
Andrew R
AR
Williams
Lynn M
LM
D020413:3' Untranslated Regions / Q000235:genetics; D001665:Binding Sites / Q000235:genetics; D002478:Cells, Cultured; D042863:Cyclin-Dependent Kinase 9 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020672:I-kappa B Proteins / Q000235:genetics / Q000378:metabolism; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000235:genetics; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000235:genetics / Q000378:metabolism; D051996:Transcription Factor RelA / Q000235:genetics; D014158:Transcription, Genetic / Q000187:drug effects; D015533:Transcriptional Activation; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D020413:3' Untranslated Regions; D020672:I-kappa B Proteins; D008070:Lipopolysaccharides; D016328:NF-kappa B; C000606204:NFKBIA protein, human; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D000072000:NF-KappaB Inhibitor alpha; D042863:Cyclin-Dependent Kinase 9; D012319:RNA Polymerase II
Nucleic Acids Res. 2004 Jun 24;32(11):e88
15247321
Infect Immun. 2000 Sep;68(9):5447-9
10948181
Proc Natl Acad Sci U S A. 2007 Oct 23;104(43):16850-5
17940009
J Biol Chem. 1998 Aug 14;273(33):21178-86
9694874
J Immunol. 2002 May 15;168(10):4837-45
11994432
Eur J Immunol. 1995 Sep;25(9):2668-72
7589143
N Engl J Med. 2009 Nov 19;361(21):2033-45
19890111
J Leukoc Biol. 2005 Jan;77(1):3-15
15522916
J Biol Chem. 1995 Apr 21;270(16):9558-63
7721885
Genes Dev. 2005 May 1;19(9):1116-27
15879558
J Immunol. 2002 Sep 1;169(5):2253-63
12193690
Cell. 2009 Jul 10;138(1):114-28
19596239
Science. 1998 Sep 4;281(5382):1509-12
9727977
Cold Spring Harb Perspect Biol. 2009 Oct;1(4):a000224
20066094
Nat Genet. 2008 May;40(5):553-9
18443593
Nat Med. 2003 Oct;9(10):1245-50
14520364
Nat Rev Immunol. 2010 Mar;10(3):170-81
20154735
Mol Cell. 2001 Aug;8(2):327-37
11545735
J Immunol. 2009 Jul 15;183(2):1197-206
19542371
J Biol Chem. 1999 Nov 5;274(45):31868-74
10542212
Immunology. 2004 Nov;113(3):281-92
15500614
Proc Natl Acad Sci U S A. 2005 Jun 14;102(24):8686-91
15937121
J Biol Chem. 2001 Aug 24;276(34):31793-9
11431468
Cell. 2009 Jul 10;138(1):129-45
19596240
Mol Cell Biol. 2000 Apr;20(8):2629-34
10733565
EMBO J. 2001 Jul 16;20(14):3760-70
11447117
Blood. 2010 Jun 3;115(22):4421-30
20237317
Immunity. 1999 Jan;10(1):39-49
10023769
Nat Immunol. 2003 Jun;4(6):551-6
12754507
Annu Rev Immunol. 2001;19:683-765
11244051
Cell. 1993 Oct 22;75(2):263-74
8402911
Mol Cell Biol. 2009 Mar;29(5):1375-87
19103749
Proc Natl Acad Sci U S A. 2009 Oct 27;106(43):18207-12
19820169
Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):8211-5
9653166
J Immunol. 2004 Jan 1;172(1):567-76
14688368