Involvement of the Pro-oncoprotein TLS (Translocated in Liposarcoma) in Nuclear Factor-κB p65-mediated Transcription as a Coactivator

Abstract

In this study, we have demonstrated that translocated in liposarcoma (TLS), also termed FUS, is an interacting molecule of the p65 (RelA) subunit of the transcription factor nuclear factor κB (NF-κB) using a yeast two-hybrid screen. We confirmed the interaction between TLS and p65 by the pull-down assay in vitro and by a coimmunoprecipitation experiment followed by Western blot of the cultured cell in vivo. TLS was originally identified as part of a fusion protein with CHOP arising from chromosomal translocation in human myxoid liposarcomas. TLS has been shown to be involved in TFIID complex formation and associated with RNA polymerase II. However, the role of TLS in transcriptional regulation has not yet been clearly elucidated. We found that TLS enhanced the NF-κB-mediated transactivation induced by physiological stimuli such as tumor necrosis factor α, interleukin-1β, and overexpression of NF-κB-inducing kinase. TLS augmented NF-κB-dependent promoter activity of the intercellular adhesion molecule-1 gene and interferon-β gene. These results suggest that TLS acts as a coactivator of NF-κB and plays a pivotal role in the NF-κB-mediated transactivation. In this study, we have demonstrated that translocated in liposarcoma (TLS), also termed FUS, is an interacting molecule of the p65 (RelA) subunit of the transcription factor nuclear factor κB (NF-κB) using a yeast two-hybrid screen. We confirmed the interaction between TLS and p65 by the pull-down assay in vitro and by a coimmunoprecipitation experiment followed by Western blot of the cultured cell in vivo. TLS was originally identified as part of a fusion protein with CHOP arising from chromosomal translocation in human myxoid liposarcomas. TLS has been shown to be involved in TFIID complex formation and associated with RNA polymerase II. However, the role of TLS in transcriptional regulation has not yet been clearly elucidated. We found that TLS enhanced the NF-κB-mediated transactivation induced by physiological stimuli such as tumor necrosis factor α, interleukin-1β, and overexpression of NF-κB-inducing kinase. TLS augmented NF-κB-dependent promoter activity of the intercellular adhesion molecule-1 gene and interferon-β gene. These results suggest that TLS acts as a coactivator of NF-κB and plays a pivotal role in the NF-κB-mediated transactivation. Nuclear factor κB (NF-κB)1 is an inducible cellular transcription factor that regulates a wide variety of cellular and viral genes including cytokines, cell adhesion molecules and human immunodeficiency virus (1Baeuerle P.A. Baichwal V.R. Adv. Immunol. 1997; 65: 111-137Crossref PubMed Google Scholar, 2Baldwin Jr., A.S. Annu. Rev. Immunol. 1996; 14: 649-683Crossref PubMed Scopus (5552) Google Scholar, 3Hayashi T. Ueno Y. Okamoto T. J. Biol. Chem. 1993; 268: 11380-11388Abstract Full Text PDF PubMed Google Scholar, 4Okamoto T. Sakurada S. Yang J.P. Merin J.P. Curr. Top. Cell Regul. 1997; 35: 149-161Crossref PubMed Scopus (75) Google Scholar, 5Ghosh S. May M.J. Kopp E.B. Annu. Rev. Immunol. 1998; 16: 225-260Crossref PubMed Scopus (4585) Google Scholar). The members of the NF-κB family in mammalian cells include the proto-oncogene c-Rel, RelA (p65), RelB, NFkB1 (p50/105), and NFkB2 (p52/p100). These proteins share a conserved 300-amino acid region known as the Rel homology domain, which is responsible for DNA binding, dimerization, and nuclear translocation of NF-κB (1Baeuerle P.A. Baichwal V.R. Adv. Immunol. 1997; 65: 111-137Crossref PubMed Google Scholar, 2Baldwin Jr., A.S. Annu. Rev. Immunol. 1996; 14: 649-683Crossref PubMed Scopus (5552) Google Scholar, 4Okamoto T. Sakurada S. Yang J.P. Merin J.P. Curr. Top. Cell Regul. 1997; 35: 149-161Crossref PubMed Scopus (75) Google Scholar, 5Ghosh S. May M.J. Kopp E.B. Annu. Rev. Immunol. 1998; 16: 225-260Crossref PubMed Scopus (4585) Google Scholar). In most cells, Rel family members form hetero- and homodimers with distinct specificities in various combinations. p65, RelB, and c-Rel are transcriptionally active members of the NF-κB family, whereas p50 and p52 primarily serve as DNA binding subunits (1Baeuerle P.A. Baichwal V.R. Adv. Immunol. 1997; 65: 111-137Crossref PubMed Google Scholar, 2Baldwin Jr., A.S. Annu. Rev. Immunol. 1996; 14: 649-683Crossref PubMed Scopus (5552) Google Scholar, 4Okamoto T. Sakurada S. Yang J.P. Merin J.P. Curr. Top. Cell Regul. 1997; 35: 149-161Crossref PubMed Scopus (75) Google Scholar, 5Ghosh S. May M.J. Kopp E.B. Annu. Rev. Immunol. 1998; 16: 225-260Crossref PubMed Scopus (4585) Google Scholar). These proteins play fundamental roles in immune and inflammatory responses and in the control of cell proliferation (4Okamoto T. Sakurada S. Yang J.P. Merin J.P. Curr. Top. Cell Regul. 1997; 35: 149-161Crossref PubMed Scopus (75) Google Scholar,6Kajino S. Suganuma M. Teranishi F. Takahashi N. Tetsuka T. Ohara H. Itoh M. Okamoto T. Oncogene. 2000; 19: 2233-2239Crossref PubMed Scopus (34) Google Scholar, 7Tetsuka T. Uranishi H. Imai H. Ono T. Sonta S. Takahashi N. Asamitsu K. Okamoto T. J. Biol. Chem. 2000; 275: 4383-4390Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar, 8Yang J.P. Hori M. Sanda T. Okamoto T. J. Biol. Chem. 1999; 274: 15662-15670Abstract Full Text Full Text PDF PubMed Scopus (141) Google Scholar, 9Yang J.P. Hori M. Takahashi N. Kawabe T. Kato H. Okamoto T. Oncogene. 1999; 18: 5177-5186Crossref PubMed Scopus (104) Google Scholar). A common feature of the regulation of NF-κB is the sequestration in the cytoplasm as an inactive complex with a class of inhibitory molecules known as IκBs (2Baldwin Jr., A.S. Annu. Rev. Immunol. 1996; 14: 649-683Crossref PubMed Scopus (5552) Google Scholar, 10Sun S.C. Ganchi P.A. Beraud C. Ballard D.W. Greene W.C. Proc. Natl. Acad. Sci. U. S. A. 1994; 91: 1346-1350Crossref PubMed Scopus (162) Google Scholar). Treatment of cells with a variety of inducers such as phorbol esters, interleukin-1 (IL-1), and tumor necrosis factor α (TNF-α) results in phosphorylation, ubiquitination, and degradation of the IκB proteins (5Ghosh S. May M.J. Kopp E.B. Annu. Rev. Immunol. 1998; 16: 225-260Crossref PubMed Scopus (4585) Google Scholar, 11Mercurio F. Manning A.M. Curr. Opin. Cell Biol. 1999; 11: 226-232Crossref PubMed Scopus (446) Google Scholar, 12Zandi E. Karin M. Mol. Cell. Biol. 1999; 19: 4547-4551Crossref PubMed Scopus (303) Google Scholar). The degradation of IκB proteins exposes the nuclear localization sequence in the remaining NF-κB dimers, followed by the rapid translocation of NF-κB to the nucleus where it activates the target genes by binding to the DNA regulatory element (1Baeuerle P.A. Baichwal V.R. Adv. Immunol. 1997; 65: 111-137Crossref PubMed Google Scholar, 2Baldwin Jr., A.S. Annu. Rev. Immunol. 1996; 14: 649-683Crossref PubMed Scopus (5552) Google Scholar, 4Okamoto T. Sakurada S. Yang J.P. Merin J.P. Curr. Top. Cell Regul. 1997; 35: 149-161Crossref PubMed Scopus (75) Google Scholar, 5Ghosh S. May M.J. Kopp E.B. Annu. Rev. Immunol. 1998; 16: 225-260Crossref PubMed Scopus (4585) Google Scholar). The protein regions responsible for the transcriptional activation (called “transactivation domain”) of p65, RelB, and c-Rel have been mapped in their unique C-terminal regions. p65 contains at least two independent transactivation domains within its C-terminal 120 amino acids (Fig. 1 A) (13Moore P.A. Ruben S.M. Rosen C.A. Mol. Cell. Biol. 1993; 13: 1666-1674Crossref PubMed Google Scholar, 14Schmitz M.L. Baeuerle P.A. EMBO J. 1991; 10: 3805-3817Crossref PubMed Scopus (664) Google Scholar, 15Schmitz 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 (138) Google Scholar, 16Schmitz M.L. dos Santos Silva M.A. Baeuerle P.A. J. Biol. Chem. 1995; 270: 15576-15584Crossref PubMed Scopus (144) Google Scholar). One of these transactivation domains, TA1, is confined to the C-terminal 30 amino acids of p65. The second transactivation domain, TA2, is located within the N-terminally adjacent 90 amino acids and contains TA1-like domain and leucine-rich regions. Since the nuclear translocation and DNA binding of NF-κB were not sufficient for gene induction (17Yoza B.K. Hu J.Y.Q. McCall C.E. J. Biol. Chem. 1996; 271: 18306-18309Abstract Full Text Full Text PDF PubMed Scopus (66) Google Scholar, 18Bergmann M. Hart L. Lindsay M. Barnes P.J. Newton R. J. Biol. Chem. 1998; 273: 6607-6610Abstract Full Text Full Text PDF PubMed Scopus (167) Google Scholar), it was suggested that interactions with other protein molecules through the transactivation domain (15Schmitz 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 (138) Google Scholar, 19Perkins N.D. Felzien L.K. Betts J.C. Leung K. Beach D.H. Nabel G.J. Science. 1997; 275: 523-527Crossref PubMed Scopus (666) Google Scholar, 20Gerritsen M.E. Williams A.J. Neish A.S. Moore S. Shi Y. Collins T. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 2927-2932Crossref PubMed Scopus (711) Google Scholar) as well as its modification by phosphorylation (16Schmitz M.L. dos Santos Silva M.A. Baeuerle P.A. J. Biol. Chem. 1995; 270: 15576-15584Crossref PubMed Scopus (144) Google Scholar) might play a critical role. It has been previously reported that transcriptional activation of NF-κB requires multiple coactivator proteins including CREB-binding protein (CBP)/p300 (19Perkins N.D. Felzien L.K. Betts J.C. Leung K. Beach D.H. Nabel G.J. Science. 1997; 275: 523-527Crossref PubMed Scopus (666) Google Scholar, 20Gerritsen M.E. Williams A.J. Neish A.S. Moore S. Shi Y. Collins T. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 2927-2932Crossref PubMed Scopus (711) Google Scholar), CBP-associated factor, and steroid receptor coactivator 1 (21Sheppard K.A. Rose D.W. Haque Z.K. Kurokawa R. McInerney E. Westin S. Thanos Collins T. Mol. Cell. Biol. 1999; 19: PubMed Google Scholar). These have activity to the and also to the transcriptional p65 was also found to with a identified coactivator which transcriptional activation by vitro A.M. P.A. S. S. R. 1999; PubMed Scopus Google Scholar). In to the transcriptional activation of be by transcription through interaction with the activation In the of the of p65 with transcription such as and has been demonstrated (15Schmitz 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 (138) Google Scholar, C. H. E. Y. C. Mol. Cell. Biol. 1993; 13: PubMed Scopus Google Scholar, Mol. Cell. Biol. 1994; 14: PubMed Scopus Google Scholar, M.J. A.J. 1993; PubMed Scopus Google Scholar, A. R. EMBO J. 1998; PubMed Scopus Google Scholar, A. S. R. J. Biol. Chem. 2000; 275: Full Text Full Text PDF PubMed Scopus Google Scholar). It is that interactions with NF-κB its transcriptional of the NF-κB transcriptional activity is by interaction with and and its is by interaction with and at multiple In yeast two-hybrid two of was found to with the region of p65 (RelA) and DNA binding in the to the of IκBs J.P. Hori M. Sanda T. Okamoto T. J. Biol. Chem. 1999; 274: 15662-15670Abstract Full Text Full Text PDF PubMed Scopus (141) Google Scholar). The other proteins found to with p65 to the family, including of and of T. Uranishi H. Imai H. Ono T. Sonta S. Takahashi N. Asamitsu K. Okamoto T. J. Biol. Chem. 2000; 275: 4383-4390Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar), previously known as nuclear S.M. 1998; 14: Full Text Full Text PDF PubMed Scopus Google Scholar, M. 1998; PubMed Scopus Google Scholar). in liposarcoma (TLS), also known as FUS, was originally identified through its fusion to a of the protein family of transcription in human myxoid liposarcoma with the chromosomal translocation A. N. 1993; PubMed Scopus Google Scholar, A. R. 1993; PubMed Scopus Google Scholar). TLS has homology to protein the family A. T. J. M. L. Mol. Cell. Biol. 1998; 18: PubMed Scopus Google These genes were found to be involved in through chromosomal translocation with other genes of transcription the region of these proteins a transcriptional domain, and the of proteins a domain these fusion proteins active for their transcriptional M. L. Oncogene. 1994; Google Scholar, A. L. Oncogene. 1999; 18: PubMed Scopus Google Scholar, R. J. F. M. G. J. Mol. Cell. Biol. 1994; 14: PubMed Scopus Google Scholar). The C-terminal of TLS the sequence domain is by and is associated with the fusion of the to the domain of a transcription factor A. N. 1993; PubMed Scopus Google Scholar, A. R. 1993; PubMed Scopus Google Scholar, H. K. Y. M. 1994; Google Scholar). TLS was shown to with a of the TFIID complex in cells A. T. J. M. L. Mol. Cell. Biol. 1998; 18: PubMed Scopus Google Scholar, A. Y. L. EMBO J. 1996; PubMed Scopus Google Scholar). a of was with RNA polymerase at the active H. Mol. Cell. Biol. 1995; PubMed Scopus Google Scholar). In TLS was shown to be associated with RNA polymerase through its domain L. Mol. Cell. Biol. 2000; PubMed Scopus Google Scholar). In this study, we that TLS with NF-κB p65 through the C-terminal transactivation domain and activates NF-κB-mediated The yeast two-hybrid interaction assay that the region between two transactivation domains of p65, TA1-like and TA1, is for the interaction with We confirmed the interaction between p65 and TLS in vitro using the fusion proteins and in blot In TLS transactivation and NF-κB-dependent gene These that TLS the transcriptional activity of to and were from T. S. T. of of L. A. and J. S. of and T. and have been previously T. Uranishi H. Imai H. Ono T. Sonta S. Takahashi N. Asamitsu K. Okamoto T. J. Biol. Chem. 2000; 275: 4383-4390Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar). the mammalian for the TLS was from with the and and in at to form a form of the TLS was by polymerase using as a with the These were and in at the to form which the fusion protein of and p65, the of human p65 acids was by polymerase using as a with was and in the of and p65 from with and were in the of of a of the human immunodeficiency sequence of promoter has been previously T. Asamitsu K. Yang J.P. Takahashi N. Tetsuka T. A. A. Okamoto T. 1998; 14: PubMed Scopus Google Scholar). The other was from contains of the binding of the The yeast two-hybrid was as previously T. Uranishi H. Imai H. Ono T. Sonta S. Takahashi N. Asamitsu K. Okamoto T. J. Biol. Chem. 2000; 275: 4383-4390Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar, 8Yang J.P. Hori M. Sanda T. Okamoto T. J. Biol. Chem. 1999; 274: 15662-15670Abstract Full Text Full Text PDF PubMed Scopus (141) Google Scholar, 9Yang J.P. Hori M. Takahashi N. Kawabe T. Kato H. Okamoto T. Oncogene. 1999; 18: 5177-5186Crossref PubMed Scopus (104) Google Scholar). The various of the p65 C-terminal regions to amino acids and were to DNA binding domain using the were for activation of was as a for it in the was with and the human was to the transactivation domain in the were for the to the with and were from that were for activity and identified by and their amino acid were with and for of the interacting cells were in with and were using to the the cells were and the were for activity was by the as previously J.P. Hori M. Sanda T. Okamoto T. J. Biol. Chem. 1999; 274: 15662-15670Abstract Full Text Full Text PDF PubMed Scopus (141) Google Scholar). was by activity using the as an The are as the in to control of independent and were from of fusion proteins and which and the TLS was by polymerase using as a with These were with and in at fusion proteins were in and as J.P. Hori M. Sanda T. Okamoto T. J. Biol. Chem. 1999; 274: 15662-15670Abstract Full Text Full Text PDF PubMed Scopus (141) Google Scholar). In vitro interaction were as previously T. Uranishi H. Imai H. Ono T. Sonta S. Takahashi N. Asamitsu K. Okamoto T. J. Biol. Chem. 2000; 275: 4383-4390Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar, 8Yang J.P. Hori M. Sanda T. Okamoto T. J. Biol. Chem. 1999; 274: 15662-15670Abstract Full Text Full Text PDF PubMed Scopus (141) Google Scholar). p65 and proteins were with by the in using the to the of fusion proteins were of and two with 1 of 1 The were in of the and were with the proteins for at with The were with 1 of and two with 1 of proteins were with 30 of for and by of cells were cultured for and with 1 The was with 1 of control and control at of protein were and the was for 1 The were with 1 of were by in were by and The was with and proteins were by enhanced as previously T. J. 1996; PubMed Scopus Google Scholar, T. J. Biol. Chem. 1996; 271: Full Text Full Text PDF PubMed Scopus Google Scholar). to human TLS was a from K. and M. was by with acids the of p65 by the was proteins interacting with the p65 subunit of we the yeast two-hybrid using the unique C-terminal region of NF-κB p65 as a and in 1 various of p65 amino acids and were to domain in the these was as a for the it in assay was to human to the transcriptional activation domain in the yeast 90 and in a from the was with the yeast to the DNA and with and the gene for TLS in to and the IκB family including which were previously shown to with two-hybrid interaction between p65 and cells were with various proteins to and is a that TLS to acids to were and to for at 30 least of were for activity using a assay for activity activity activity not in a cells were with various proteins to and is a that TLS to acids to were and to for at 30 least of were for activity using a assay for activity activity activity not the region of p65 involved in the binding to various regions of the protein were to in the and with TLS to transcriptional activation were by activity and by of yeast cells with and and (Fig. 1 and not in the in the and were shown to be for the interaction with and These results that the region of p65 responsible for the interaction with TLS within the amino acid sequence It was previously that the of TLS might as a transcriptional in fusion with other transcription such as CHOP and M. L. Oncogene. 1994; Google Scholar, H. R. 1994; PubMed Scopus Google Scholar), transcriptional activity of the TLS its has not been It is suggested that TLS transcriptional activation by with transcription such as and we TLS acts as a transcriptional coactivator to the of a this we various fusion proteins of TLS and domain TLS its C-terminal demonstrated in of these fusion proteins transcriptional activation from the the control of it was suggested that TLS as a coactivator of transcription it was to the of the the interaction between TLS and p65, we vitro interaction assay using various TLS proteins in fusion with The p65 protein was by in in the of and was with fusion proteins shown in p65 to A binding between p65 and was also In this assay control for interaction with also to p65. However, p65 binding was with A coimmunoprecipitation experiment was to TLS with p65 in cultured cells were with and p65 and Cell from these cells were with (Fig. its C-terminal The were by and with TLS and whereas control not and These results demonstrated that TLS with the p65 subunit of NF-κB in as well as in We the of TLS NF-κB-dependent gene In the of TLS was gene from the by of with of in TLS augmented the NF-κB-mediated transactivation in a the was gene from by with the of gene activation was to by the of However, was the transcription a control in which κB were was the activation by the of of was These of TLS were not the of an p65 Western blot of the cell in the protein of p65 (Fig. TLS augmented gene from human promoter binding for NF-κB not the of TLS the transactivation domain of p65 and its we for fusion proteins in which the domain of was with various of p65 The of of transactivation of these and by TLS were in TLS augmented the transactivation by and by and In was of TLS the of and These that the of TLS transactivation to the C-terminal domain of p65 and were for the physiological of the interaction between TLS and p65, we have the of TLS the human intercellular adhesion molecule-1 promoter an NF-κB binding T. M.A. Neish A.S. Thanos T. J. 1995; PubMed Scopus Google Scholar, J. Biol. Chem. 2000; 275: Full Text Full Text PDF PubMed Scopus Google Scholar). In to with p65 of physiological stimuli such as were also of the were cells with and the of and were demonstrated in TLS augmented the gene in a (Fig. TLS enhanced the gene induced by and it that the of TLS gene is through we have confirmed this by using an the NF-κB and the shown in TLS augmented NF-κB-dependent gene from induced by and by NF-κB-inducing an involved in the NF-κB activation by a the κB was activation by was and of of TLS was demonstrated not These demonstrated that the of TLS were also the NF-κB-dependent gene the interaction of TLS with p65 the cell was from cells that were with TLS control in the of The cell was with and the proteins were to a and by demonstrated in TLS was with p65 in cells by A of TLS was with p65 cells were not by coimmunoprecipitation was control was In not the protein of p65 and TLS as demonstrated by using These results that TLS with p65 physiological for the activation of NF-κB subunit p65 contains at least two independent transactivation domains, and TA2, located in the C-terminal contains TA1-like domain, regions are for its a demonstrated that the region between the TA1-like domain of and the domain acids of was for the activity of not sufficient for the transcriptional activity (16Schmitz M.L. dos Santos Silva M.A. Baeuerle P.A. J. Biol. Chem. 1995; 270: 15576-15584Crossref PubMed Scopus (144) Google Scholar). the C-terminal of p65 as a in the yeast two-hybrid we have identified TLS as an interacting We have the region of p65 acids for the interaction with TLS (Fig. have that TLS acts as a of the NF-κB transactivation and TLS a common feature with a of including and These proteins have been found associated with TFIID A. T. J. M. L. Mol. Cell. Biol. 1998; 18: PubMed Scopus Google Scholar, A. Y. L. EMBO J. 1996; PubMed Scopus Google Scholar) and are in transcriptional activation M. L. Oncogene. 1994; Google Scholar, A. L. Oncogene. 1999; 18: PubMed Scopus Google Scholar, H. R. 1994; PubMed Scopus Google Scholar, H. K. R. M. Mol. Cell. Biol. 1999; 19: PubMed Scopus Google Scholar). TFIID is a complex of and a of T. R. Cell. 1991; Full Text PDF PubMed Scopus Google Scholar, P.A. J. Biol. Chem. 1993; 268: Full Text PDF PubMed Google Scholar, J.C. L. 1994; 523-527Crossref PubMed Scopus Google Scholar, N. 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In this study, we also that TLS was with TFIID as well as with p65 in cultured cells not These suggest that TLS gene through between p65 and transcriptional TLS was originally identified as a fusion protein with CHOP in human liposarcoma and found in other such as in which the of TLS was to the C-terminal region of A. N. 1993; PubMed Scopus Google Scholar, A. R. 1993; PubMed Scopus Google Scholar, H. K. Y. M. 1994; Google Scholar). The fusion with the of TLS was suggested to be a for these transcriptional to have the by their by the target gene H. R. 1994; PubMed Scopus Google Scholar, H. K. R. M. Mol. Cell. Biol. 1999; 19: PubMed Scopus Google Scholar). In demonstrated that the of TLS a transcriptional activity to (Fig. The C-terminal region of which is by the domain of transcription through chromosomal also a transcriptional activity by fusion with it was suggested that the of chromosomal translocation TLS might also the transcriptional results that TLS acts as a transcriptional were suggested by other TLS to RNA in a vitro and in cells A. N. 1993; PubMed Scopus Google Scholar, M. L. Oncogene. 1994; Google Scholar, H. Y. Oncogene. 1997; 14: PubMed Scopus Google Scholar) and in rapid H. R. 1994; PubMed Scopus Google Scholar, H. Y. Oncogene. 1997; 14: PubMed Scopus Google Scholar). These with the that TLS with a of the TFIID complex in cells A. T. J. M. L. Mol. Cell. Biol. 1998; 18: PubMed Scopus Google Scholar, A. Y. L. EMBO J. 1996; PubMed Scopus Google Scholar), suggest that TLS in transcriptional regulation and by in nuclear formation C. G. M. 1995; Scholar). we previously demonstrated that the region acids within the p65 transactivation domain with T. Uranishi H. Imai H. Ono T. Sonta S. Takahashi N. Asamitsu K. Okamoto T. J. Biol. Chem. 2000; 275: 4383-4390Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar). We found that of transactivation was by and not are to the binding of TLS and with p65, it is that the transcriptional activity of NF-κB is through the binding of interacting proteins with such as TLS and In these suggest a of TLS in regulation of gene The of NF-κB to with in to other transcription including (15Schmitz 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 (138) Google Scholar), C. H. E. Y. C. Mol. Cell. Biol. 1993; 13: PubMed Scopus Google Scholar, M.J. A.J. 1993; PubMed Scopus Google Scholar), A. R. EMBO J. 1998; PubMed Scopus Google Scholar, A. S. R. J. Biol. Chem. 2000; 275: Full Text Full Text PDF PubMed Scopus Google Scholar), and (19Perkins N.D. Felzien L.K. Betts J.C. Leung K. Beach D.H. Nabel G.J. Science. 1997; 275: 523-527Crossref PubMed Scopus (666) Google Scholar, 20Gerritsen M.E. Williams A.J. Neish A.S. Moore S. Shi Y. Collins T. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 2927-2932Crossref PubMed Scopus (711) Google Scholar), be to its transcriptional activity as well as its to various transcriptional such as of acid and J. J. Biol. Chem. 2000; 275: Full Text Full Text PDF PubMed Scopus Google Scholar) and T. Uranishi H. Imai H. Ono T. Sonta S. Takahashi N. Asamitsu K. Okamoto T. J. Biol. Chem. 2000; 275: 4383-4390Abstract Full Text Full Text PDF PubMed Scopus (78) Google Scholar). We T. S. T. L. A. J. S. and T. for We K. and M. for the of nuclear factor κB of protein CREB-binding protein domain transcriptional activation domain intercellular adhesion molecule-1 NF-κB-inducing nuclear localization of translocated in liposarcoma tumor necrosis factor