An Acute Phase Response Factor/NF-B Site Downstream of the junB Gene That Mediates Responsiveness to Interleukin-6 in a Murine Plasmacytoma (*)

The immediate early gene, junB, is induced by interleukin-6 (IL-6) in plasmacytomas. In order to identify enhancers that mediate this effect, we cloned upstream and downstream sequences flanking the gene into a luciferase reporter gene vector containing the junB promoter and evaluated the IL-6 inducibility of these sequences by transient expression in an IL-6-dependent plasmacytoma cell line. Although a 6.5-kilobase fragment of upstream flanking sequence did not increase the IL-6 inducibility of the junB promoter, a 222-base pair fragment was identified in 2.1 kilobases of downstream flanking sequence that both up-regulates the promoter and confers inducibility by IL-6. Point mutation of an acute phase response factor (APRF) site within this region significantly reduced up-regulation of the promoter in cells grown continuously in IL-6, as well as inducibility upon restimulation of cells with IL-6 after withdrawal from the growth factor. Point mutation of an NF-κB site sharing five nucleotides with the APRF site reduced up-regulation of the promoter but not inducibility by IL-6, whereas mutation of two other NF-κB sites in the 222-base pair fragment had no effect on expression. Western blotting of nuclear proteins purified by DNA affinity chromatography revealed inducible binding of Stat3 and constitutive binding of NF-κB p65 to the APRF/NF-κB site.

[1]  M. Potter,et al.  Induction of Plasma-Cell Neoplasms in Strain BALB/c Mice with Mineral Oil and Mineral Oil Adjuvants , 1962, Nature.

[2]  M. Potter,et al.  Induction of Plasma Cell Tumours in BALB/c Mice with 2,6,10,14-Tetramethylpentadecane (Pristane) , 1969, Nature.

[3]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[4]  M. Hayashi,et al.  Template activities of the phi X-174 replicative allomorphic deoxyribonucleic acids. , 1971, Biochemistry.

[5]  H. Echols,et al.  Effect of Circularity and Superhelicity on Transcription from Bacteriophage λ DNA , 1973 .

[6]  L. Lau,et al.  Identification of a set of genes expressed during the G0/G1 transition of cultured mouse cells. , 1985, The EMBO journal.

[7]  R. Nordan,et al.  A macrophage-derived factor required by plasmacytomas for survival and proliferation in vitro. , 1986, Science.

[8]  L. Lau,et al.  Expression of a set of growth-related immediate early genes in BALB/c 3T3 cells: coordinate regulation with c-fos or c-myc. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[9]  G. Nabel,et al.  An inducible transcription factor activates expression of human immunodeficiency virus in T cells , 1987, Nature.

[10]  R. Nordan,et al.  Regulation of murine plasmacytoma transferrin receptor expression and G1 traversal by plasmacytoma cell growthfactor , 1988, Journal of cellular physiology.

[11]  D. Nathans,et al.  Induction of protooncogene c-jun by serum growth factors. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[12]  R. Moritz,et al.  Characterization of a recombinant murine interleukin-6: assignment of disulfide bonds. , 1988, Biochemical and biophysical research communications.

[13]  L. Lau,et al.  A gene activated by growth factors is related to the oncogene v-jun. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[14]  T. Komori,et al.  Pathogenic significance of interleukin-6 (IL-6/BSF-2) in Castleman's disease , 1989 .

[15]  B. Klein,et al.  Interleukin-6 is a potent myeloma-cell growth factor in patients with aggressive multiple myeloma. , 1989, Blood.

[16]  B. Wasylyk,et al.  The c-ets proto-oncogenes encode transcription factors that cooperate with c-Fos and c-Jun for transcriptional activation , 1990, Nature.

[17]  R. Wall,et al.  Interleukin-6 signals activating junB and TIS11 gene transcription in a B-cell hybridoma , 1991, Molecular and cellular biology.

[18]  Ian Magrath,et al.  A chemiluminescent assay for quantitation of β-galactosidase in the femtogram range: Application to quantitation of β-galactosidase in lacZ-transfected cells , 1991 .

[19]  V. N. Roa,et al.  Ets-related protein Elk-1 is homologous to the c-fos regulatory factor p62TCF , 1991, Nature.

[20]  J. Auwerx,et al.  Activation of junB by PKC and PKA signal transduction through a novel cis-acting element. , 1991, Nucleic acids research.

[21]  R. Jove,et al.  Regulation of the junB gene by v-src , 1992, Molecular and cellular biology.

[22]  P. Sharp,et al.  The p50 subunit of NF-kappa B associates with the NF-IL6 transcription factor. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[23]  Dimitris Thanos,et al.  The High Mobility Group protein HMG I(Y) is required for NF-κB-dependent virus induction of the human IFN-β gene , 1992, Cell.

[24]  R. Tantravahi,et al.  Myb and Ets proteins cooperate in transcriptional activation of the mim-1 promoter. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[25]  R. Treisman,et al.  Characterization of SAP-1, a protein recruited by serum response factor to the c-fos serum response element , 1992, Cell.

[26]  K. Yokoyama,et al.  Two cis-regulatory elements that mediate different signaling pathways for serum-dependent activation of the junB gene. , 1993, The Journal of biological chemistry.

[27]  G. Nabel,et al.  A cooperative interaction between NF‐kappa B and Sp1 is required for HIV‐1 enhancer activation. , 1993, The EMBO journal.

[28]  L. Sanders,et al.  Transcriptional regulatory elements downstream of the JunB gene. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[29]  Qiang Zhou,et al.  Association between proto-oncoprotein Rel and TATA-binding protein mediates transcriptional activation by NF-κB , 1993, Nature.

[30]  T. Hirano,et al.  Identification of a novel interleukin-6 response element containing an Ets-binding site and a CRE-like site in the junB promoter , 1993, Molecular and cellular biology.

[31]  W. Greene,et al.  Cross‐coupling of the NF‐kappa B p65 and Fos/Jun transcription factors produces potentiated biological function. , 1993, The EMBO journal.

[32]  S. Akira,et al.  Transcription factors NF-IL6 and NF-kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Darnell,et al.  Stat3: a STAT family member activated by tyrosine phosphorylation in response to epidermal growth factor and interleukin-6. , 1994, Science.

[34]  T. Hirano,et al.  Transcriptional activation of the IL-6 response element in the junB promoter is mediated by multiple Stat family proteins. , 1994, Biochemical and biophysical research communications.

[35]  A. Ray,et al.  Physical association and functional antagonism between the p65 subunit of transcription factor NF-kappa B and the glucocorticoid receptor. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[36]  H. Baumann,et al.  Separate Signaling Mechanisms Are Involved in the Control of STAT Protein Activation and Gene Regulation via the Interleukin 6 Response Element by the Box 3 Motif of gp130 (*) , 1995, The Journal of Biological Chemistry.

[37]  J. Blenis,et al.  Requirement of serine phosphorylation for formation of STAT-promoter complexes. , 1995, Science.

[38]  P. Coffer,et al.  Transcriptional regulation of the junB promoter: analysis of STAT-mediated signal transduction. , 1995, Oncogene.