A Constitutive Active MEK → ERK Pathway Negatively Regulates NF-κB-dependent Gene Expression by Modulating TATA-binding Protein Phosphorylation

Endotoxin-induced cytokine gene expression is regulated, in part, by NF-κB. We have shown that both the ERK and p38 mitogen-activated protein (MAP) kinases are necessary for cytokine gene transcription and that the p38 MAP kinase is required for NF-κB-driven transcription, so we hypothesized that the MEK → ERK pathway regulated NF-κB-driven transcription as well. We found that a constitutive active MEK → ERK pathway inhibited NF-κB-driven transcription. In addition, both PD 98059 and a dominant negative ERK2 augmented NF-κB-driven transcription; however, neither PD 98059 nor MEK1 altered NF-κB activation at any level. The constitutive active MEK → ERK pathway inhibited the phosphorylation of TBP, which is necessary for both interaction with RelA and binding to the TATA box. Due to the fact that we have shown that the p38 MAP kinase modulates TBP activation, we evaluated the effect of the constitutive active MEK → ERK pathway on p38 MAP kinase activity. We found that the MEK → ERK pathway negatively regulates NF-κB-driven transcription, in part, by inhibiting p38 MAP kinase activity. Thus, the ERK and p38 MAP kinases have differential effects on NF-κB-driven transcription. Endotoxin-induced cytokine gene expression is regulated, in part, by NF-κB. We have shown that both the ERK and p38 mitogen-activated protein (MAP) kinases are necessary for cytokine gene transcription and that the p38 MAP kinase is required for NF-κB-driven transcription, so we hypothesized that the MEK → ERK pathway regulated NF-κB-driven transcription as well. We found that a constitutive active MEK → ERK pathway inhibited NF-κB-driven transcription. In addition, both PD 98059 and a dominant negative ERK2 augmented NF-κB-driven transcription; however, neither PD 98059 nor MEK1 altered NF-κB activation at any level. The constitutive active MEK → ERK pathway inhibited the phosphorylation of TBP, which is necessary for both interaction with RelA and binding to the TATA box. Due to the fact that we have shown that the p38 MAP kinase modulates TBP activation, we evaluated the effect of the constitutive active MEK → ERK pathway on p38 MAP kinase activity. We found that the MEK → ERK pathway negatively regulates NF-κB-driven transcription, in part, by inhibiting p38 MAP kinase activity. Thus, the ERK and p38 MAP kinases have differential effects on NF-κB-driven transcription. nuclear factor κB IκB kinase transcription factor IIB TATA-binding protein transcription factor IID mitogen-activated protein extracellular signal-regulated kinase c-Jun NH2-terminal MAP kinase MAP kinase kinase MAP kinase kinase kinase 1 activating protein-1 CCAAT-enhancer binding protein β activating transcription factor-2 interleukin lipopolysaccharide chloramphenicol acetyltransferase polyacrylamide gel electrophoresis tumor necrosis factor A transcription factor that is necessary for the transcription of many genes involved in inflammatory and immune responses is nuclear factor κB (NF-κB)1 (1Baeuerle P.A. Henkel T. Annu. Rev. Immunol. 1994; 12: 141-179Crossref PubMed Scopus (4641) Google Scholar,2Thanos D. Maniatis T. Cell. 1995; 80: 529-532Abstract Full Text PDF PubMed Scopus (1221) Google Scholar). NF-κB is a dimeric protein that is composed of members of the Rel family of transcription factors. The regulation of NF-κB-dependent gene expression can occur at multiple levels after cell stimulation. NF-κB is held in the cytoplasm by an inhibitor protein, IκB, in quiescent cells (3Finco T.S. Baldwin A.S. Immunity. 1995; 3: 263-272Abstract Full Text PDF PubMed Scopus (385) Google Scholar, 4Matthews J.R. Hay R.T. Int. J. Biochem. Cell Biol. 1995; 27: 865-879Crossref PubMed Scopus (72) Google Scholar). In stimulated cells, the phosphorylation of IκBα on Ser-32 and Ser-36 and IκBβ on Ser-19 and Ser-23 by IKKα and IKKβ, respectively, results in their subsequent degradation (3Finco T.S. Baldwin A.S. Immunity. 1995; 3: 263-272Abstract Full Text PDF PubMed Scopus (385) Google Scholar, 5Baldwin Jr., A.S. Annu. Rev. Immunol. 1996; 14: 649-683Crossref PubMed Scopus (5645) Google Scholar, 6Brockman J.A. Scherer D.C. McKinsey T.A. Hall S.M. Qi X. Lee W.Y. Ballard D.W. Mol. Cell. Biol. 1995; 15: 2809-2818Crossref PubMed Google Scholar, 7DiDonato J. Mercurio F. Rosette C. Wu-Li J. Suyang H. Ghosh S. Karin M. Mol. Cell. Biol. 1996; 16: 1295-1304Crossref PubMed Google Scholar, 8Chen Z.J. Parent L. Maniatis T. Cell. 1996; 84: 853-862Abstract Full Text Full Text PDF PubMed Scopus (878) Google Scholar). IκB degradation allows NF-κB to translocate to the nucleus and bind to DNA (7DiDonato J. Mercurio F. Rosette C. Wu-Li J. Suyang H. Ghosh S. Karin M. Mol. Cell. Biol. 1996; 16: 1295-1304Crossref PubMed Google Scholar,9Whiteside S.T. Ernst M.K. LeBail O. Laurent-Winter C. Rice N. Israel A. Mol. Cell. Biol. 1995; 15: 5339-5345Crossref PubMed Google Scholar, 10Traenckner E.B. Pahl H.L. Henkel T. Schmidt K.N. Wilk S. Baeuerle P.A. EMBO J. 1995; 14: 2876-2883Crossref PubMed Scopus (937) Google Scholar, 11O'Connell M.A. Bennett B.L. Mercurio F. Manning A.M. Mackman N. J. Biol. Chem. 1998; 273: 30410-30414Abstract Full Text Full Text PDF PubMed Scopus (164) Google Scholar). Transcriptional activity, however, also requires the phosphorylation of the transcriptionally active NF-κB subunits, RelA (p65), c-Rel, and RelB. In fact, several studies have shown that the phosphorylation of p65 is necessary for transcription to be initiated (12Hayashi T. Sekine T. Okamoto T. J. Biol. Chem. 1993; 268: 26790-26795Abstract Full Text PDF PubMed Google Scholar, 13Naumann M. Scheidereit C. EMBO J. 1994; 13: 4597-4607Crossref PubMed Scopus (329) Google Scholar, 14Schmitz M.L. dos Santos Silva M.A. Baeuerle P.A. J. Biol. Chem. 1995; 270: 15576-15584Abstract Full Text Full Text PDF PubMed Scopus (144) Google Scholar, 15Zhong H. SuYang H. Erdjument-Bromage H. Tempst P. Ghosh S. Cell. 1997; 89: 413-424Abstract Full Text Full Text PDF PubMed Scopus (729) Google Scholar, 16Zhong H. Voll R.E. Ghosh S. Mol. Cell. 1998; 1: 661-671Abstract Full Text Full Text PDF PubMed Scopus (1033) Google Scholar). The interaction of the p65 subunit with the basal transcription factors IIB (TFIIB) and TATA-binding protein (TBP) is also necessary for NF-κB-driven transcription (17Kerr L.D. Ransone L.J. Wamsley P. Schmitt M.J. Boyer T.G. Zhou Q. Berk A.J. Verma I.M. Nature. 1993; 365: 412-419Crossref PubMed Scopus (134) Google Scholar, 18Xu X. Prorock C. Ishikawa H. Maldonado E. Ito Y. Gelinas C. Mol. Cell. Biol. 1993; 13: 6733-6741Crossref PubMed Scopus (98) Google Scholar, 19Blair W.S. Bogerd H.P. Madore S.J. Cullen B.R. Mol. Cell. Biol. 1994; 14: 7226-7234Crossref PubMed Scopus (104) Google Scholar, 20Schmitz M.L. Stelzer G. Altmann H. Meisterernst M. Baeuerle P.A. J. Biol. Chem. 1995; 270: 7219-7226Abstract Full Text Full Text PDF PubMed Scopus (139) Google Scholar, 21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar). The phosphorylation of TBP is required for both its interaction with the p65 subunit and its binding to the TATA box (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar, 22Chibazakura T. Watanabe F. Kitajima S. Tsukada K. Yasukochi Y. Teraoka H. Eur. J. Biochem. 1997; 247: 1166-1173Crossref PubMed Scopus (41) Google Scholar). The mitogen-activated protein (MAP) kinases are a family of second messenger kinases that is essential for transferring signals from the cell surface to the nucleus. We have shown that the p38 MAP kinase regulates NF-κB-dependent gene expression by modulating the phosphorylation and subsequent activation of TBP (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar). The regulation of NF-κB-dependent transcription is important due to the fact that most cytokine promoter sequences have active NF-κB sites and thus suggests the mechanism by which the p38 MAP kinase controls cytokine gene expression. Other MAP kinases have been linked to cytokine gene expression. In addition to others (23Reimann T. Buscher D. Hipskind R.A. Krautwald S. Lohmann-Matthes M.L. Baccarini M. J. Immunol. 1994; 153: 5740-5749PubMed Google Scholar, 24Carter A.B. Monick M.M. Hunninghake G.W. Am. J. Respir. Cell. Mol. Biol. 1999; 20: 751-758Crossref PubMed Scopus (284) Google Scholar), we have shown that activation of the ERK kinase is important for the induction of cytokine genes in macrophages exposed to endotoxin (LPS). The MEK → ERK kinase pathway has been shown to be required for the activation of AP-1 and C/EBPβ, both of which are transcription factors involved in the transcription of cytokine genes (25Nakajima T. Kinoshita S. Sasagawa T. Sasaki K. Naruto M. Kishimoto T. Akira S. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 2207-2211Crossref PubMed Scopus (521) Google Scholar, 26Minden A. Lin A. Smeal T. Derijard B. Cobb M. Davis R. Karin M. Mol. Cell. Biol. 1994; 14: 6683-6688Crossref PubMed Scopus (440) Google Scholar, 27Frost J.A. Geppert T.D. Cobb M.H. Feramisco J.R. Proc. Natl. Acad. Sci. U. S. A. 1994; 91: 3844-3848Crossref PubMed Scopus (205) Google Scholar, 28Frost J.A. Alberts A.S. Sontag E. Guan K. Mumby M.C. Feramisco J.R. Mol. Cell. Biol. 1994; 14: 6244-6252Crossref PubMed Scopus (93) Google Scholar, 29DeSilva D.R. Feeser W.S. Tancula E.J. Scherle P.A. J. Exp. Med. 1996; 183: 2017-2023Crossref PubMed Scopus (113) Google Scholar). Another study has shown that sustained activation of the MEK → ERK pathway resulted in induction of IL-5, IL-6, IL-10, and IL-13 genes in T cells (30Chen D. Heath V. O'Garra A. Johnston J. McMahon M. J. Immunol. 1999; 163: 5796-5805PubMed Google Scholar). None of these studies, however, evaluated the effect of the MEK → ERK pathway on NF-κB-dependent transcription. Based on our previous data showing that the ERK kinase regulated IL-6 transcription (24Carter A.B. Monick M.M. Hunninghake G.W. Am. J. Respir. Cell. Mol. Biol. 1999; 20: 751-758Crossref PubMed Scopus (284) Google Scholar) and that the p38 MAP kinase was necessary for NF-κB-dependent gene expression (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar), we analyzed the role of the MEK → ERK pathway in regulating NF-κB-dependent transcription. a promoter by we found that a constitutive active MEK1 expression inhibited NF-κB-dependent in cells, and PD a inhibitor of NF-κB activation was altered by the MEK → ERK In the constitutive active MEK1 expression inhibited the phosphorylation of a protein, which be to with the p65 subunit and to bind to the TATA box. In addition, the constitutive active MEK → ERK pathway the of the p38 MAP that the ERK and p38 MAP kinases have differential effects on NF-κB-dependent gene expression. The cell was from The cells in with and in phosphorylation studies the cells in with the and for in kinase the cells in with and NF-κB-dependent gene expression was evaluated a by of the in a The from and from have been C. Maniatis T. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, N. Y. K. J. Biol. Chem. Full Text PDF PubMed Google Scholar). The from of and have been (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar, M. Davis J. Biol. Chem. 1994; Full Text PDF PubMed Google Scholar). The was by of the of the p38 MAP kinase and a expression The and from the to the after the cells stimulated with at a of activity, which was to protein, was after which was to be the of activity. acetyltransferase which to protein, after as G.W. Monick M.M. B. J. PubMed Google Scholar). the MEK kinase PD 98059 at was 1 to with A NF-κB was with as A.B. Monick M.M. Hunninghake G.W. Am. J. Respir. Cell. Mol. Biol. 1998; PubMed Scopus Google Scholar, M.M. A.B. Hunninghake G.W. J. Biol. Chem. 1999; 274: Full Text Full Text PDF PubMed Scopus Google Scholar), and the on a polyacrylamide cell as (24Carter A.B. Monick M.M. Hunninghake G.W. Am. J. Respir. Cell. Mol. Biol. 1999; 20: 751-758Crossref PubMed Scopus (284) Google Scholar, M.M. A.B. G. R. Hunninghake G.W. J. Immunol. 1999; Google Scholar). The p38 ERK2 MAP kinases from the at with a p38 ERK2 respectively, to with In kinase was as (TBP) as a (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar, 24Carter A.B. Monick M.M. Hunninghake G.W. Am. J. Respir. Cell. Mol. Biol. 1999; 20: 751-758Crossref PubMed Scopus (284) Google Scholar, M.M. A.B. G. R. Hunninghake G.W. J. Immunol. 1999; Google Scholar). of the with the was to kinase in the cells stimulated with and activity, which was to protein, was after which was to be the of activity. to as (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar). ERK2 and p38 MAP kinase at and the was at The and the the sequences and respectively, from the in the and the and at a cells with an the and the cells as (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar). The the the from cell with the to with from to at The by and phosphorylation was by which to protein, are shown as with the a with the of to be to the role of the ERK kinase in regulating NF-κB-dependent gene transcription, we that the ERK kinase and that PD 98059 was in that the ERK cells stimulated with at and a was for We found that was at was as as and to levels at of ERK2 MAP kinase of the In c-Jun as a and results found of ERK kinase have in cells with the ERK2 kinase data that the ERK2 kinase in The MEK PD has been shown to be an and inhibitor of L. S.J. A.J. Proc. Natl. Acad. Sci. U. S. A. 1995; PubMed Scopus Google Scholar, D.R. A. P. J. Biol. Chem. 1995; 270: Full Text Full Text PDF PubMed Scopus Google Scholar, C. R.A. K. M.A. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar). In to that PD 98059 was in cells, we both in kinase to activation of the ERK2 the p38 MAP We a with the and in cells and in the of PD was to levels 1 results for and in ERK2 in c-Jun as the In p38 MAP kinase as by phosphorylation of and PD 98059 effect on kinase activity. PD 98059 in cells exposed to studies that PD 98059 is and for of the MEK → ERK pathway with PD 98059 p38 MAP as by kinase the role of the MEK → ERK pathway in regulating NF-κB-dependent transcription, we NF-κB-dependent promoter the We the kinase to that was active in with the and an the constitutive active MEK1 expression and MEK1 and in cells MEK1 and stimulated with the was at a ERK2 in kinase c-Jun as the results data that ERK2 both in the constitutive active MEK1 expression we evaluated its role in regulating NF-κB-driven gene transcription. In these studies, we the with an the MEK1 expression in cells the in cells the constitutive active MEK1 was to levels A was with cells in the of PD 98059 after the PD 98059 augmented in cells the and in cells the constitutive active MEK1 PD 98059 the effect of MEK1 on NF-κB-dependent in a we the with an the dominant negative The results from these the results found with PD in cells with the and was augmented in cells the dominant negative ERK2 MAP kinase these results that the MEK → ERK pathway negatively regulates NF-κB-dependent gene expression. In to the role of the MEK → ERK kinase pathway in the regulation NF-κB-driven transcription, we evaluated p65 phosphorylation was by MEK1 ERK2 activity. cells in the of PD 98059 for 1 and stimulated with and nuclear protein was that NF-κB and DNA of the MEK → ERK pathway with PD 98059 effect on and binding the constitutive active MEK1 altered p65 cells with the and an the constitutive active MEK1 expression the cells in and stimulated with was to phosphorylation in cells the and the constitutive active MEK1 expression effect on phosphorylation of of the an these studies that the MEK → ERK pathway the activation of NF-κB by its and DNA binding the phosphorylation of the p65 and of pathway with PD 98059 the of NF-κB. Due to the fact that NF-κB-driven transcription requires activation of TBP and activation is on the p38 MAP kinase (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar), we evaluated the negative regulation of NF-κB-dependent transcription by the MEK → ERK pathway was to of TBP We the with an the constitutive active MEK1 expression The cells in and stimulated with was from cell to phosphorylation of in cells the In phosphorylation was in cells MEK1 of of the suggests that the MEK → ERK pathway negatively regulates NF-κB-dependent gene transcription, in part, by modulating the activation of the of inhibiting NF-κB by the constitutive active MEK1 expression we a gene by a an both of which have cells with an and in cells the In cells the constitutive active MEK1 a as by in A and In cells with the a in activity, and was by in cells the constitutive active MEK1 data the MEK → ERK pathway negatively regulates gene expression that requires NF-κB by modulating TBP the data that cytokine gene expression is of NF-κB the MEK → ERK pathway is necessary for the activation of AP-1 and both of which have binding sites the and Due to the fact that the p38 MAP kinase is a (21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar) and the constitutive active MEK1 is a negative of NF-κB-dependent transcription by activation of TBP, we evaluated a constitutive active MEK → ERK pathway inhibited p38 MAP kinase activity. the constitutive active MEK1 expression inhibited the p38 MAP we in p38 MAP kinase cells with an the constitutive active MEK1 expression The p38 MAP kinase was from cell and kinase was (TBP) as the (TBP) was in by p38 MAP kinase from cells with the and was in cells MEK1 for p38 MAP kinase of the The phosphorylation of and is required for p38 MAP kinase activation J. S. M. J. Davis J. Biol. Chem. 1995; 270: Full Text Full Text PDF PubMed Scopus Google Scholar), so we the constitutive active MEK1 expression inhibited the phosphorylation of a p38 MAP kinase In these we the with an the MEK1 expression The cells in to with The protein was to the phosphorylation of and phosphorylation was in cells MEK1 for the protein of the an these studies that the p38 MAP kinase (TBP) and a constitutive active MEK → ERK pathway p38 MAP kinase both in and in by its studies also of a mechanism by which the MEK → ERK at in part, negatively regulates NF-κB-dependent gene expression. NF-κB-dependent gene expression can be regulated at multiple levels after cell stimulation. The regulation occur in the cytoplasm NF-κB is held by the inhibitor protein, studies have shown that an of both the and p38 MAP kinase both IKKα and IKKβ, and in with the kinase to the S. J.A. Lin A. Mol. Cell. Biol. 1998; PubMed Google Scholar, M.J. Y. S. Mercurio F. M. Cobb M.H. Cell. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar). In addition, the of dominant of inhibited NF-κB-dependent gene expression by S. J.A. Lin A. Mol. Cell. Biol. 1998; PubMed Google Scholar, M.J. Y. S. Mercurio F. M. Cobb M.H. Cell. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar, M.A. S. J. Davis T. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar). of the results in subsequent IκB and (3Finco T.S. Baldwin A.S. Immunity. 1995; 3: 263-272Abstract Full Text PDF PubMed Scopus (385) Google J.A. Scherer D.C. McKinsey T.A. Hall S.M. Qi X. Lee W.Y. Ballard D.W. Mol. Cell. Biol. 1995; 15: 2809-2818Crossref PubMed Google Scholar, 7DiDonato J. Mercurio F. Rosette C. Wu-Li J. Suyang H. Ghosh S. Karin M. Mol. Cell. Biol. 1996; 16: 1295-1304Crossref PubMed Google Scholar, S.T. Ernst M.K. LeBail O. Laurent-Winter C. Rice N. Israel A. Mol. Cell. Biol. 1995; 15: 5339-5345Crossref PubMed Google Scholar, 10Traenckner E.B. Pahl H.L. Henkel T. Schmidt K.N. Wilk S. Baeuerle P.A. EMBO J. 1995; 14: 2876-2883Crossref PubMed Scopus (937) Google Scholar). The of NF-κB is also on the phosphorylation of the p65 subunit (12Hayashi T. Sekine T. Okamoto T. J. Biol. Chem. 1993; 268: 26790-26795Abstract Full Text PDF PubMed Google Scholar, 13Naumann M. Scheidereit C. EMBO J. 1994; 13: 4597-4607Crossref PubMed Scopus (329) Google Scholar, 14Schmitz M.L. dos Santos Silva M.A. Baeuerle P.A. J. Biol. Chem. 1995; 270: 15576-15584Abstract Full Text Full Text PDF PubMed Scopus (144) Google Scholar, 15Zhong H. SuYang H. Erdjument-Bromage H. Tempst P. Ghosh S. Cell. 1997; 89: 413-424Abstract Full Text Full Text PDF PubMed Scopus (729) Google Scholar, 16Zhong H. Voll R.E. Ghosh S. Mol. Cell. 1998; 1: 661-671Abstract Full Text Full Text PDF PubMed Scopus (1033) Google Scholar). data that the regulation by the MEK → ERK pathway of activation, IκB NF-κB and phosphorylation of the p65 NF-κB are and the protein is to as the interaction with transcription occur to transcription. of NF-κB have been shown to with both transcription as C/EBPβ, and M.A. P.A. Proc. Natl. Acad. Sci. U. S. A. 89: PubMed Scopus Google Scholar, B. Baldwin Jr., A.S. Mol. Cell. Biol. 1993; 13: PubMed Google Scholar, B. Baldwin Jr., A.S. Ballard D.W. P. P. EMBO J. 1993; 12: PubMed Scopus Google Scholar, A.B. E. Mol. Cell. Biol. 1994; 14: PubMed Scopus Google Scholar, R. P. A.M. T. J. Mol. Cell. Biol. 1993; 13: PubMed Google Scholar, D. Maniatis T. Cell. 1993; Full Text PDF PubMed Scopus Google Scholar), and basal transcription as and TBP (17Kerr L.D. Ransone L.J. Wamsley P. Schmitt M.J. Boyer T.G. Zhou Q. Berk A.J. Verma I.M. Nature. 1993; 365: 412-419Crossref PubMed Scopus (134) Google Scholar, 18Xu X. Prorock C. Ishikawa H. Maldonado E. Ito Y. Gelinas C. Mol. Cell. Biol. 1993; 13: 6733-6741Crossref PubMed Scopus (98) Google Scholar, 19Blair W.S. Bogerd H.P. Madore S.J. Cullen B.R. Mol. Cell. Biol. 1994; 14: 7226-7234Crossref PubMed Scopus (104) Google Scholar, 20Schmitz M.L. Stelzer G. Altmann H. Meisterernst M. Baeuerle P.A. J. Biol. Chem. 1995; 270: 7219-7226Abstract Full Text Full Text PDF PubMed Scopus (139) Google Scholar). The MEK → ERK pathway is to of the transcription factors that with NF-κB (25Nakajima T. Kinoshita S. Sasagawa T. Sasaki K. Naruto M. Kishimoto T. Akira S. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 2207-2211Crossref PubMed Scopus (521) Google Scholar, 26Minden A. Lin A. Smeal T. Derijard B. Cobb M. Davis R. Karin M. Mol. Cell. Biol. 1994; 14: 6683-6688Crossref PubMed Scopus (440) Google Scholar, 27Frost J.A. Geppert T.D. Cobb M.H. Feramisco J.R. Proc. Natl. Acad. Sci. U. S. A. 1994; 91: 3844-3848Crossref PubMed Scopus (205) Google Scholar, 28Frost J.A. Alberts A.S. Sontag E. Guan K. Mumby M.C. Feramisco J.R. Mol. Cell. Biol. 1994; 14: 6244-6252Crossref PubMed Scopus (93) Google Scholar, 29DeSilva D.R. Feeser W.S. Tancula E.J. Scherle P.A. J. Exp. Med. 1996; 183: 2017-2023Crossref PubMed Scopus (113) Google Scholar), has been shown to activation of In addition to others M.L. Stelzer G. Altmann H. Meisterernst M. Baeuerle P.A. J. Biol. Chem. 1995; 270: 7219-7226Abstract Full Text Full Text PDF PubMed Scopus (139) Google Scholar, 21Carter A.B. Knudtson K.L. Monick M.M. Hunninghake G.W. J. Biol. Chem. 1999; 274: 30858-30863Abstract Full Text Full Text PDF PubMed Scopus (424) Google Scholar), we have shown that the interaction of TBP with the p65 subunit of NF-κB is required for we that the p38 MAP kinase TBP, modulating its In the we that the interaction of TBP and the p65 subunit is necessary for NF-κB-dependent transcription. The however, is that of the MEK → ERK pathway the phosphorylation of TBP and thus negatively regulates NF-κB activity. The negative regulation of NF-κB-dependent gene expression by the constitutive active MEK1 expression that the ERK and p38 MAP kinases NF-κB activity. Other studies have shown that these MAP kinases have effects on gene expression. activation of the p38 and MAP kinases and of the ERK MAP kinase was found to be for in cells stimulated with factor M. J. Davis 1995; 270: PubMed Scopus Google Scholar). In the p38 MAP kinase was shown to of the ERK MAP kinase inhibited transcription in macrophages stimulated with M.M. J. Immunol. 1999; 163: Google Scholar). In addition to effects on gene these kinases have also been shown to negatively activation of the p38 MAP kinase with in cells inhibited the activation of the ERK2 MAP kinase C. R.A. K. M.A. J. Biol. 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Thus, of the MEK PD activation of the mechanism by which both PD 98059 and the dominant negative ERK2 expression NF-κB-dependent gene expression. data that the constitutive activation of the MEK → ERK pathway negatively regulates NF-κB-dependent transcription of TBP In addition, is the study showing that a constitutive active MEK → ERK pathway p38 MAP kinase and the and required for NF-κB-dependent gene expression. be of in studies to the constitutive active MEK1 expression constitutive activation of a We the of DNA for the in and Davis for their and P. for