Abstract
To date, three functional domains have been defined in c-Fos and v-Fos proteins and have been shown to play a role in transactivation: the leucine zipper mediating hetero-dimerization, the basic DNA contact site, and a C-terminally located transactivation domain (C-TA) harbouring the HOB1 and HOB2 motifs. While the bZip region, consisting of the leucine zipper and the DNA contact site, is indispensable for transformation, the C-TA domain is not required and is actually altered by internal deletions in the FBR-MuSV. We now show that the N-terminal regions of c-Fos and v-Fos contain a second transactivation domain (N-TA). A functionally crucial motif within the N-TA domain, termed NTM, was pinpointed to a approximately 25 amino acid stretch around positions 60-84 which is highly conserved in FosB. Analysis of LexA fusion proteins showed that the N-TA domains of both c-Fos and FosB function in an autonomous fashion in both fibroblasts and yeast. Most importantly, deletion of the NTM motif impairs the transforming properties of v-Fos. Apart from the bZip region, the N-TA domain is the only functional domain required for transformation by v-Fos, at least when its expression is driven by the strong FBR-MuSV-LTR promoter.
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- Abate C., Marshak D. R., Curran T. Fos is phosphorylated by p34cdc2, cAMP-dependent protein kinase and protein kinase C at multiple sites clustered within regulatory regions. Oncogene. 1991 Dec;6(12):2179–2185. [PubMed] [Google Scholar]
- Abken H., Reifenrath B. A procedure to standardize CAT reporter gene assay. Nucleic Acids Res. 1992 Jul 11;20(13):3527–3527. doi: 10.1093/nar/20.13.3527. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Angel P., Imagawa M., Chiu R., Stein B., Imbra R. J., Rahmsdorf H. J., Jonat C., Herrlich P., Karin M. Phorbol ester-inducible genes contain a common cis element recognized by a TPA-modulated trans-acting factor. Cell. 1987 Jun 19;49(6):729–739. doi: 10.1016/0092-8674(87)90611-8. [DOI] [PubMed] [Google Scholar]
- Baichwal V. R., Tjian R. Control of c-Jun activity by interaction of a cell-specific inhibitor with regulatory domain delta: differences between v- and c-Jun. Cell. 1990 Nov 16;63(4):815–825. doi: 10.1016/0092-8674(90)90147-7. [DOI] [PubMed] [Google Scholar]
- Boyle W. J., Smeal T., Defize L. H., Angel P., Woodgett J. R., Karin M., Hunter T. Activation of protein kinase C decreases phosphorylation of c-Jun at sites that negatively regulate its DNA-binding activity. Cell. 1991 Feb 8;64(3):573–584. doi: 10.1016/0092-8674(91)90241-p. [DOI] [PubMed] [Google Scholar]
- Cohen D. R., Curran T. The structure and function of the fos proto-oncogene. Crit Rev Oncog. 1989;1(1):65–88. [PubMed] [Google Scholar]
- Cohen D. R., Curran T. fra-1: a serum-inducible, cellular immediate-early gene that encodes a fos-related antigen. Mol Cell Biol. 1988 May;8(5):2063–2069. doi: 10.1128/mcb.8.5.2063. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Curran T., Verma I. M. FBR murine osteosarcoma virus. I. Molecular analysis and characterization of a 75,000-Da gag-fos fusion product. Virology. 1984 May;135(1):218–228. doi: 10.1016/0042-6822(84)90132-6. [DOI] [PubMed] [Google Scholar]
- Gentz R., Rauscher F. J., 3rd, Abate C., Curran T. Parallel association of Fos and Jun leucine zippers juxtaposes DNA binding domains. Science. 1989 Mar 31;243(4899):1695–1699. doi: 10.1126/science.2494702. [DOI] [PubMed] [Google Scholar]
- Gius D., Cao X. M., Rauscher F. J., 3rd, Cohen D. R., Curran T., Sukhatme V. P. Transcriptional activation and repression by Fos are independent functions: the C terminus represses immediate-early gene expression via CArG elements. Mol Cell Biol. 1990 Aug;10(8):4243–4255. doi: 10.1128/mcb.10.8.4243. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Horii T., Ogawa T., Ogawa H. Nucleotide sequence of the lexA gene of E. coli. Cell. 1981 Mar;23(3):689–697. doi: 10.1016/0092-8674(81)90432-3. [DOI] [PubMed] [Google Scholar]
- Jenuwein T., Müller D., Curran T., Müller R. Extended life span and tumorigenicity of nonestablished mouse connective tissue cells transformed by the fos oncogene of FBR-MuSV. Cell. 1985 Jun;41(2):629–637. doi: 10.1016/s0092-8674(85)80035-0. [DOI] [PubMed] [Google Scholar]
- Jenuwein T., Müller R. Structure-function analysis of fos protein: a single amino acid change activates the immortalizing potential of v-fos. Cell. 1987 Feb 27;48(4):647–657. doi: 10.1016/0092-8674(87)90243-1. [DOI] [PubMed] [Google Scholar]
- Jooss K., Müller R. Analysis of temperature-sensitive functions of Fos: lack of a correlation between transformation and TRE-dependent trans-activation. Oncogene. 1992 Oct;7(10):1933–1939. [PubMed] [Google Scholar]
- Kouzarides T., Ziff E. The role of the leucine zipper in the fos-jun interaction. Nature. 1988 Dec 15;336(6200):646–651. doi: 10.1038/336646a0. [DOI] [PubMed] [Google Scholar]
- Kovary K., Rizzo C. A., Ryseck R. P., Noguchi T., Raynoschek C., Pelosin J. M., Bravo R. Constitutive expression of FosB and its short form, FosB/SF, induces malignant cell transformation in rat-1A cells. New Biol. 1991 Sep;3(9):870–879. [PubMed] [Google Scholar]
- Lassar A. B., Thayer M. J., Overell R. W., Weintraub H. Transformation by activated ras or fos prevents myogenesis by inhibiting expression of MyoD1. Cell. 1989 Aug 25;58(4):659–667. doi: 10.1016/0092-8674(89)90101-3. [DOI] [PubMed] [Google Scholar]
- Lech K., Anderson K., Brent R. DNA-bound Fos proteins activate transcription in yeast. Cell. 1988 Jan 29;52(2):179–184. doi: 10.1016/0092-8674(88)90506-5. [DOI] [PubMed] [Google Scholar]
- Lin A., Frost J., Deng T., Smeal T., al-Alawi N., Kikkawa U., Hunter T., Brenner D., Karin M. Casein kinase II is a negative regulator of c-Jun DNA binding and AP-1 activity. Cell. 1992 Sep 4;70(5):777–789. doi: 10.1016/0092-8674(92)90311-y. [DOI] [PubMed] [Google Scholar]
- Lucibello F. C., Lowag C., Neuberg M., Müller R. trans-repression of the mouse c-fos promoter: a novel mechanism of Fos-mediated trans-regulation. Cell. 1989 Dec 22;59(6):999–1007. doi: 10.1016/0092-8674(89)90756-3. [DOI] [PubMed] [Google Scholar]
- Lucibello F. C., Müller R. Proto-oncogenes encoding transcriptional regulators: unravelling the mechanisms of oncogenic conversion. Crit Rev Oncog. 1991;2(4):259–276. [PubMed] [Google Scholar]
- Lucibello F. C., Neuberg M., Jenuwein T., Müller R. Multiple regions of v-Fos protein involved in the activation of AP1-dependent transcription: is trans-activation crucial for transformation? New Biol. 1991 Jul;3(7):671–677. [PubMed] [Google Scholar]
- Lucibello F. C., Slater E. P., Jooss K. U., Beato M., Müller R. Mutual transrepression of Fos and the glucocorticoid receptor: involvement of a functional domain in Fos which is absent in FosB. EMBO J. 1990 Sep;9(9):2827–2834. doi: 10.1002/j.1460-2075.1990.tb07471.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsui M., Tokuhara M., Konuma Y., Nomura N., Ishizaki R. Isolation of human fos-related genes and their expression during monocyte-macrophage differentiation. Oncogene. 1990 Mar;5(3):249–255. [PubMed] [Google Scholar]
- Mumberg D., Lucibello F. C., Schuermann M., Müller R. Alternative splicing of fosB transcripts results in differentially expressed mRNAs encoding functionally antagonistic proteins. Genes Dev. 1991 Jul;5(7):1212–1223. doi: 10.1101/gad.5.7.1212. [DOI] [PubMed] [Google Scholar]
- Müller R. Cellular and viral fos genes: structure, regulation of expression and biological properties of their encoded products. Biochim Biophys Acta. 1986;823(3):207–225. doi: 10.1016/0304-419x(86)90003-x. [DOI] [PubMed] [Google Scholar]
- Müller R., Müller D. Co-transfection of normal NIH/3T3 DNA and retroval LTR sequences: a novel strategy for the detection of potential c-onc genes. EMBO J. 1984 May;3(5):1121–1127. doi: 10.1002/j.1460-2075.1984.tb01939.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nakabeppu Y., Nathans D. A naturally occurring truncated form of FosB that inhibits Fos/Jun transcriptional activity. Cell. 1991 Feb 22;64(4):751–759. doi: 10.1016/0092-8674(91)90504-r. [DOI] [PubMed] [Google Scholar]
- Neuberg M., Schuermann M., Hunter J. B., Müller R. Two functionally different regions in Fos are required for the sequence-specific DNA interaction of the Fos/Jun protein complex. Nature. 1989 Apr 13;338(6216):589–590. doi: 10.1038/338589a0. [DOI] [PubMed] [Google Scholar]
- Neuberg M., Schuermann M., Müller R. Mutagenesis of the DNA contact site in Fos protein: compatibility with the scissors grip model and requirement for transformation. Oncogene. 1991 Aug;6(8):1325–1333. [PubMed] [Google Scholar]
- Nishina H., Sato H., Suzuki T., Sato M., Iba H. Isolation and characterization of fra-2, an additional member of the fos gene family. Proc Natl Acad Sci U S A. 1990 May;87(9):3619–3623. doi: 10.1073/pnas.87.9.3619. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oliviero S., Robinson G. S., Struhl K., Spiegelman B. M. Yeast GCN4 as a probe for oncogenesis by AP-1 transcription factors: transcriptional activation through AP-1 sites is not sufficient for cellular transformation. Genes Dev. 1992 Sep;6(9):1799–1809. doi: 10.1101/gad.6.9.1799. [DOI] [PubMed] [Google Scholar]
- Pothier F., Ouellet M., Julien J. P., Guérin S. L. An improved CAT assay for promoter analysis in either transgenic mice or tissue culture cells. DNA Cell Biol. 1992 Jan-Feb;11(1):83–90. doi: 10.1089/dna.1992.11.83. [DOI] [PubMed] [Google Scholar]
- Pulverer B. J., Kyriakis J. M., Avruch J., Nikolakaki E., Woodgett J. R. Phosphorylation of c-jun mediated by MAP kinases. Nature. 1991 Oct 17;353(6345):670–674. doi: 10.1038/353670a0. [DOI] [PubMed] [Google Scholar]
- Rüther U., Komitowski D., Schubert F. R., Wagner E. F. c-fos expression induces bone tumors in transgenic mice. Oncogene. 1989 Jul;4(7):861–865. [PubMed] [Google Scholar]
- Schuermann M. An expression vector system for stable expression of oncogenes. Nucleic Acids Res. 1990 Aug 25;18(16):4945–4946. doi: 10.1093/nar/18.16.4945. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schuermann M., Hennig G., Müller R. Transcriptional activation and transformation by chimaeric Fos-estrogen receptor proteins: altered properties as a consequence of gene fusion. Oncogene. 1993 Oct;8(10):2781–2790. [PubMed] [Google Scholar]
- Schuermann M., Jooss K., Müller R. fosB is a transforming gene encoding a transcriptional activator. Oncogene. 1991 Apr;6(4):567–576. [PubMed] [Google Scholar]
- Schuermann M., Neuberg M., Hunter J. B., Jenuwein T., Ryseck R. P., Bravo R., Müller R. The leucine repeat motif in Fos protein mediates complex formation with Jun/AP-1 and is required for transformation. Cell. 1989 Feb 10;56(3):507–516. doi: 10.1016/0092-8674(89)90253-5. [DOI] [PubMed] [Google Scholar]
- Schütte J., Viallet J., Nau M., Segal S., Fedorko J., Minna J. jun-B inhibits and c-fos stimulates the transforming and trans-activating activities of c-jun. Cell. 1989 Dec 22;59(6):987–997. doi: 10.1016/0092-8674(89)90755-1. [DOI] [PubMed] [Google Scholar]
- Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smeal T., Binetruy B., Mercola D. A., Birrer M., Karin M. Oncogenic and transcriptional cooperation with Ha-Ras requires phosphorylation of c-Jun on serines 63 and 73. Nature. 1991 Dec 12;354(6353):494–496. doi: 10.1038/354494a0. [DOI] [PubMed] [Google Scholar]
- Stein B., Baldwin A. S., Jr, Ballard D. W., Greene W. C., Angel P., Herrlich P. Cross-coupling of the NF-kappa B p65 and Fos/Jun transcription factors produces potentiated biological function. EMBO J. 1993 Oct;12(10):3879–3891. doi: 10.1002/j.1460-2075.1993.tb06066.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sutherland J. A., Cook A., Bannister A. J., Kouzarides T. Conserved motifs in Fos and Jun define a new class of activation domain. Genes Dev. 1992 Sep;6(9):1810–1819. doi: 10.1101/gad.6.9.1810. [DOI] [PubMed] [Google Scholar]
- Turner R., Tjian R. Leucine repeats and an adjacent DNA binding domain mediate the formation of functional cFos-cJun heterodimers. Science. 1989 Mar 31;243(4899):1689–1694. doi: 10.1126/science.2494701. [DOI] [PubMed] [Google Scholar]
- Van Beveren C., Enami S., Curran T., Verma I. M. FBR murine osteosarcoma virus. II. Nucleotide sequence of the provirus reveals that the genome contains sequences acquired from two cellular genes. Virology. 1984 May;135(1):229–243. doi: 10.1016/0042-6822(84)90133-8. [DOI] [PubMed] [Google Scholar]
- Van Beveren C., van Straaten F., Curran T., Müller R., Verma I. M. Analysis of FBJ-MuSV provirus and c-fos (mouse) gene reveals that viral and cellular fos gene products have different carboxy termini. Cell. 1983 Apr;32(4):1241–1255. doi: 10.1016/0092-8674(83)90306-9. [DOI] [PubMed] [Google Scholar]
- Verma S. S., Gupta R. K., Kishore N., Sen Gupta J. A simple relationship between maximal aerobic power and body weight in Indian adolescent boys. Indian J Med Sci. 1986 Apr;40(4):93–96. [PubMed] [Google Scholar]
- Wick M., Lucibello F. C., Müller R. Inhibition of Fos- and Ras-induced transformation by mutant Fos proteins with structural alterations in functionally different domains. Oncogene. 1992 May;7(5):859–867. [PubMed] [Google Scholar]
- Wisdom R., Verma I. M. Proto-oncogene FosB: the amino terminus encodes a regulatory function required for transformation. Mol Cell Biol. 1993 May;13(5):2635–2643. doi: 10.1128/mcb.13.5.2635. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wisdom R., Yen J., Rashid D., Verma I. M. Transformation by FosB requires a trans-activation domain missing in FosB2 that can be substituted by heterologous activation domains. Genes Dev. 1992 Apr;6(4):667–675. doi: 10.1101/gad.6.4.667. [DOI] [PubMed] [Google Scholar]
- Yen J., Wisdom R. M., Tratner I., Verma I. M. An alternative spliced form of FosB is a negative regulator of transcriptional activation and transformation by Fos proteins. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5077–5081. doi: 10.1073/pnas.88.12.5077. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zerial M., Toschi L., Ryseck R. P., Schuermann M., Müller R., Bravo R. The product of a novel growth factor activated gene, fos B, interacts with JUN proteins enhancing their DNA binding activity. EMBO J. 1989 Mar;8(3):805–813. doi: 10.1002/j.1460-2075.1989.tb03441.x. [DOI] [PMC free article] [PubMed] [Google Scholar]