Home LiteratureArticle Details
PMID: 8196666 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cell transformation by c-fos requires an extended period of expression and is independent of the cell cycle.

Molecular and cellular biology ·Vol. 14 ·No. 6 ·1994-06-00 ·Pages 4295-310

Miao GG, Curran T

Abstract

The proto-oncogene transcription factors Fos and Jun form a heterodimeric complex that binds to DNA and regulates expression of specific target genes. Continuous expression of c-fos causes transformation of cultured fibroblasts and induces osteogenic sarcoma in mice. To investigate the molecular basis of fos-mediated oncogenesis, we developed a conditional cell transformation system in which Fos expression was regulated by isopropyl-beta-D-thiogalactopyranoside (IPTG). Synthesis or repression of Fos in L1-3c-fos cells occurred rapidly, within 30 min, after the removal or addition of IPTG to the culture medium. However, there was a significant delay between the induction of Fos expression and the appearance of morphological transformation. No effect was observed after 12 h of Fos expression, partial transformation was detected after 24 h, and full transformation required approximately 3 days of continuous Fos expression. Similarly, the transformed cell morphology persisted for at least 2 days after repression of Fos, and a normal phenotype was observed only after 3 days. Fos-Jun complexes, capable of binding to AP-1 sequences, were present continuously during the delay in morphological transformation. Furthermore, increased expression of several candidate Fos target genes, including those encoding Fra-1, transin (stromelysin), collagenase, and ornithine decarboxylase, was detected shortly after Fos induction. The induction of morphological transformation was not dependent on the cell cycle, as it occurred in both cycling and noncycling cells. Thus, the Fos-Jun complexes present before L1-3c-fos cells become fully transformed are transcriptionally active. These complexes disappeared, and the Fos target genes were repressed at least 2 days prior to reversion. Our results suggest that cell transformation by Fos requires increased expression of a target gene(s) with a long-lived product(s) that must reach a critical level.

Related Genes
c-fos
MeSH Terms
Animals Bone Neoplasms/genetics Cell Cycle/drug effects,physiology Cell Division/drug effects,physiology Cell Transformation, Neoplastic Cells, Cultured DNA-Binding Proteins/metabolism Fibroblasts Gene Expression/drug effects Genes, fos Isopropyl Thiogalactoside/pharmacology Kinetics Mice Osteosarcoma/genetics Proto-Oncogene Proteins c-fos/biosynthesis,metabolism Proto-Oncogene Proteins c-jun/metabolism Restriction Mapping Time Factors Transfection
Chemicals
DNA-Binding Proteins Proto-Oncogene Proteins c-fos Proto-Oncogene Proteins c-jun Isopropyl Thiogalactoside
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Miao G G
Roche Institute of Molecular Biology, Nutley, New Jersey 07110.
Curran T
References (84)
84 references, click to expand
  1. Transformation-defective, temperature-sensitive mutants of Rous sarcoma virus have a reversibly defective src-gene product.
    Cold Spring Harb Symp Quant Biol. 1980;44 Pt 2,:1007-12 PMID: 6253175
  2. Scrape-loading of Swiss 3T3 cells with ras protein rapidly activates protein kinase C in the absence of phosphoinositide hydrolysis.
    Oncogene. 1989 Jan;4(1):27-31 PMID: 2536916
  3. Extended life span and tumorigenicity of nonestablished mouse connective tissue cells transformed by the fos oncogene of FBR-MuSV.
    Cell. 1985 Jun;41(2):629-37 PMID: 2985286
  4. Stimulus-transcription coupling in the nervous system: involvement of the inducible proto-oncogenes fos and jun.
    Annu Rev Neurosci. 1991;14:421-51 PMID: 1903243
  5. Common DNA binding site for Fos protein complexes and transcription factor AP-1.
    Cell. 1988 Feb 12;52(3):471-80 PMID: 3125983
  6. Bone and haematopoietic defects in mice lacking c-fos.
    Nature. 1992 Dec 24-31;360(6406):741-5 PMID: 1465144
  7. fos-lacZ transgenic mice: mapping sites of gene induction in the central nervous system.
    Neuron. 1992 Jan;8(1):13-23 PMID: 1730004
  8. The jun and fos protein families are both required for cell cycle progression in fibroblasts.
    Mol Cell Biol. 1991 Sep;11(9):4466-72 PMID: 1908553
  9. Existence of different Fos/Jun complexes during the G0-to-G1 transition and during exponential growth in mouse fibroblasts: differential role of Fos proteins.
    Mol Cell Biol. 1992 Nov;12(11):5015-23 PMID: 1406676
  10. The mechanisms of ornithine decarboxylase deregulation in c-Ha-ras oncogene-transformed NIH 3T3 cells.
    J Biol Chem. 1988 Mar 25;263(9):4500-7 PMID: 3279036
  11. Induction of c-fos gene and protein by growth factors precedes activation of c-myc.
    Nature. 1984 Dec 20-1985 Jan 2;312(5996):716-20 PMID: 6334806
  12. A novel target cell for c-fos-induced oncogenesis: development of chondrogenic tumours in embryonic stem cell chimeras.
    EMBO J. 1991 Sep;10(9):2437-50 PMID: 1714376
  13. Leucine repeats and an adjacent DNA binding domain mediate the formation of functional cFos-cJun heterodimers.
    Science. 1989 Mar 31;243(4899):1689-94 PMID: 2494701
  14. Mutagenesis of the DNA contact site in Fos protein: compatibility with the scissors grip model and requirement for transformation.
    Oncogene. 1991 Aug;6(8):1325-33 PMID: 1909428
  15. p21H-ras-induced morphological transformation and increases in c-myc expression are independent of functional protein kinase C.
    EMBO J. 1989 Apr;8(4):1099-104 PMID: 2663469
  16. Transformation of mammalian cells by avian myelocytomatosis virus and avian erythroblastosis virus.
    Virology. 1979 Oct 30;98(2):461-5 PMID: 228482
  17. fosB is a transforming gene encoding a transcriptional activator.
    Oncogene. 1991 Apr;6(4):567-76 PMID: 1903195
  18. Growth factors regulate transin gene expression by c-fos-dependent and c-fos-independent pathways.
    Science. 1988 Dec 9;242(4884):1424-7 PMID: 2462278
  19. Identification of a small region of the v-fos gene product that is sufficient for transforming potential and growth-stimulating activity.
    Oncogene Res. 1989;5(2):79-89 PMID: 2514403
  20. Direct transcriptional stimulation of the ornithine decarboxylase gene by Fos in PC12 cells but not in fibroblasts.
    Mol Cell Biol. 1993 Aug;13(8):4657-69 PMID: 8101634
  21. Transformation of NIH 3T3 cells by microinjection of Ha-ras p21 protein.
    Nature. 1984 Aug 9-15;310(5977):508-11 PMID: 6611509
  22. Conversion of the lac repressor into an allosterically regulated transcriptional activator for mammalian cells.
    Mol Cell Biol. 1990 Jul;10(7):3343-56 PMID: 2162473
  23. Mitogenesis by v-Src: a need for active oncoprotein both in leaving G0 and in completing G1 phases of the cell cycle.
    Cell Growth Differ. 1993 Aug;4(8):671-8 PMID: 8398908
  24. Degradation of collagen in the bone-resorbing compartment underlying the osteoclast involves both cysteine-proteinases and matrix metalloproteinases.
    J Cell Physiol. 1992 Feb;150(2):221-31 PMID: 1734028
  25. Transformation by a temperature sensitive mutant of Rous sarcoma virus in the absence of serum.
    J Gen Virol. 1975 May;27(2):127-34 PMID: 167110
  26. Transformation by the fos or jun oncogene does not increase AP-1 DNA-binding activity.
    J Virol. 1993 Sep;67(9):5487-95 PMID: 8350408
  27. Pleiotropic effects of a null mutation in the c-fos proto-oncogene.
    Cell. 1992 Nov 13;71(4):577-86 PMID: 1423615
  28. Transcriptional activation and transformation by chimaeric Fos-estrogen receptor proteins: altered properties as a consequence of gene fusion.
    Oncogene. 1993 Oct;8(10):2781-90 PMID: 8378087
  29. Hormone-dependent transcriptional regulation and cellular transformation by Fos-steroid receptor fusion proteins.
    Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5114-8 PMID: 1711219
  30. Expression of c-fos in NIH3T3 cells is very low but inducible throughout the cell cycle.
    EMBO J. 1986 Apr;5(4):695-700 PMID: 3709522
  31. Molecular genetics of polyamine synthesis in eukaryotic cells.
    Trends Biochem Sci. 1990 Apr;15(4):153-8 PMID: 2187296
  32. FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA.
    J Virol. 1982 Nov;44(2):674-82 PMID: 6292525
  33. Structure-function analysis of fos protein: a single amino acid change activates the immortalizing potential of v-fos.
    Cell. 1987 Feb 27;48(4):647-57 PMID: 3028645
  34. Suppression of oncogene-induced transformation by a deletion mutant of c-jun.
    Oncogene. 1993 Apr;8(4):877-86 PMID: 8455942
  35. Transactivation of c-fos and beta-actin genes by raf as a step in early response to transmembrane signals.
    Nature. 1990 Mar 29;344(6265):463-6 PMID: 2157161
  36. Viral and cellular fos proteins: a comparative analysis.
    Cell. 1984 Feb;36(2):259-68 PMID: 6319013
  37. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene.
    Nature. 1984 Oct 4-10;311(5985):433-8 PMID: 6090941
  38. Ornithine decarboxylase activity is critical for cell transformation.
    Nature. 1992 Nov 26;360(6402):355-8 PMID: 1280331
  39. FBR murine osteosarcoma virus. I. Molecular analysis and characterization of a 75,000-Da gag-fos fusion product.
    Virology. 1984 May;135(1):218-28 PMID: 6203214
  40. The mRNA coding for the secreted protease transin is expressed more abundantly in malignant than in benign tumors.
    Proc Natl Acad Sci U S A. 1986 Dec;83(24):9413-7 PMID: 3540938
  41. Temperature-dependent alterations in sugar transport in cells infected by a temperature-sensitive mutant of Rous sarcoma virus.
    Proc Natl Acad Sci U S A. 1971 Nov;68(11):2739-41 PMID: 4330938
  42. Parallel association of Fos and Jun leucine zippers juxtaposes DNA binding domains.
    Science. 1989 Mar 31;243(4899):1695-9 PMID: 2494702
  43. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins.
    Science. 1988 Jun 24;240(4860):1759-64 PMID: 3289117
  44. Proliferative activation of quiescent Rat-1A cells by delta FosB.
    Mol Cell Biol. 1993 Jul;13(7):4157-66 PMID: 8321220
  45. Identification of a 39,000-dalton protein in cells transformed by the FBJ murine osteosarcoma virus.
    Virology. 1982 Jan 15;116(1):221-35 PMID: 6278709
  46. Expression and purification of the leucine zipper and DNA-binding domains of Fos and Jun: both Fos and Jun contact DNA directly.
    Proc Natl Acad Sci U S A. 1990 Feb;87(3):1032-6 PMID: 2105492
  47. Transformation by Fos proteins requires a C-terminal transactivation domain.
    Mol Cell Biol. 1993 Dec;13(12):7429-38 PMID: 7504176
  48. An avian transforming retrovirus isolated from a nephroblastoma that carries the fos gene as the oncogene.
    J Virol. 1987 Dec;61(12):3733-40 PMID: 2824811
  49. Microinjection of transforming ras protein induces c-fos expression.
    Mol Cell Biol. 1987 Jan;7(1):523-7 PMID: 3104768
  50. Mitogenesis of quiescent chick fibroblasts by v-Src: dependence on events at the membrane leading to early changes in AP-1.
    Oncogene. 1993 Jul;8(7):1875-86 PMID: 8510932
  51. High-efficiency transformation of mammalian cells by plasmid DNA.
    Mol Cell Biol. 1987 Aug;7(8):2745-52 PMID: 3670292
  52. Gene regulation by tyrosine kinases: src protein activates various promoters, including c-fos.
    Mol Cell Biol. 1989 Jun;9(6):2493-9 PMID: 2474755
  53. Inhibition of Fos- and Ras-induced transformation by mutant Fos proteins with structural alterations in functionally different domains.
    Oncogene. 1992 May;7(5):859-67 PMID: 1570155
  54. EGR3, a novel member of the Egr family of genes encoding immediate-early transcription factors.
    Oncogene. 1991 Jun;6(6):917-28 PMID: 1906159
  55. A cellular factor stimulates the DNA-binding activity of MyoD and E47.
    Proc Natl Acad Sci U S A. 1993 Jul 15;90(14):6483-7 PMID: 8393567
  56. Steroid-binding domains for regulating the functions of heterologous proteins in cis.
    Trends Cell Biol. 1993 Aug;3(8):278-80 PMID: 14731747
  57. Transformation suppressor activity of a Jun transcription factor lacking its activation domain.
    Nature. 1991 Aug 15;352(6336):635-8 PMID: 1907719
  58. Polyamines are essential for cell transformation by pp60v-src: delineation of molecular events relevant for the transformed phenotype.
    J Cell Biol. 1993 Aug;122(4):903-14 PMID: 7688751
  59. Analysis of inducers of the E.coli lac repressor system in mammalian cells and whole animals.
    Nucleic Acids Res. 1991 Sep 11;19(17):4647-53 PMID: 1891356
  60. Microinjection of fos-specific antibodies blocks DNA synthesis in fibroblast cells.
    Mol Cell Biol. 1988 Apr;8(4):1670-6 PMID: 2454395
  61. The role of the leucine zipper in the fos-jun interaction.
    Nature. 1988 Dec 15;336(6200):646-51 PMID: 2974122
  62. Osteoblasts are target cells for transformation in c-fos transgenic mice.
    J Cell Biol. 1993 Aug;122(3):685-701 PMID: 8335693
  63. Analysis of Fos protein complexes and Fos-related antigens by high-resolution two-dimensional gel electrophoresis.
    Oncogene. 1987 May;1(2):213-21 PMID: 3325882
  64. Redox activation of Fos-Jun DNA binding activity is mediated by a DNA repair enzyme.
    EMBO J. 1992 Sep;11(9):3323-35 PMID: 1380454
  65. Removal of a 67-base-pair sequence in the noncoding region of protooncogene fos converts it to a transforming gene.
    Proc Natl Acad Sci U S A. 1985 Aug;82(15):4987-91 PMID: 2991903
  66. Virus induction of osteosarcomas in mice.
    Science. 1966 Feb 11;151(3711):698-701 PMID: 5218049
  67. Continuous c-fos expression precedes programmed cell death in vivo.
    Nature. 1993 May 13;363(6425):166-9 PMID: 8483500
  68. A mutant p53 protein is required for maintenance of the transformed phenotype in cells transformed with p53 plus ras cDNAs.
    Proc Natl Acad Sci U S A. 1992 May 1;89(9):3952-6 PMID: 1570319
  69. Localization of pp60src within normal rat kidney cells infected with temperature-sensitive mutants (T-class) of Rous sarcoma virus.
    Cold Spring Harb Symp Quant Biol. 1980;44 Pt 2,:1013-22 PMID: 6253176
  70. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
    Anal Biochem. 1987 Apr;162(1):156-9 PMID: 2440339
  71. Candidate product of the FBJ murine osteosarcoma virus oncogene: characterization of a 55,000-dalton phosphoprotein.
    J Virol. 1982 Apr;42(1):114-22 PMID: 6283132
  72. Inducible production of c-fos antisense RNA inhibits 3T3 cell proliferation.
    Proc Natl Acad Sci U S A. 1986 Jul;83(13):4794-8 PMID: 3523478
  73. 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):507-16 PMID: 2492453
  74. Requirement for fos gene expression in the transcriptional activation of collagenase by other oncogenes and phorbol esters.
    Cell. 1988 Jul 29;54(3):325-34 PMID: 2840203
  75. Growth and differentiation of embryonic stem cells that lack an intact c-fos gene.
    Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):9306-10 PMID: 1329091
  76. How does p21ras transform cells?
    Trends Genet. 1991 Mar;7(3):91-5 PMID: 2031288
  77. Antisense RNA of proto-oncogene c-fos blocks renewed growth of quiescent 3T3 cells.
    Mol Cell Biol. 1987 Feb;7(2):639-49 PMID: 3547078
  78. ras-induced c-fos expression and proliferation in living rat fibroblasts involves C-kinase activation and the serum response element pathway.
    EMBO J. 1990 Jan;9(1):171-80 PMID: 2104799
  79. Analysis of AP-1 function in cellular transformation pathways.
    J Virol. 1994 Jun;68(6):3527-35 PMID: 8189491
  80. Platelet-derived growth factor induces rapid but transient expression of the c-fos gene and protein.
    Nature. 1984 Dec 20-1985 Jan 2;312(5996):711-6 PMID: 6514007
  81. Analysis of dimerization and DNA binding functions in Fos and Jun by domain-swapping: involvement of residues outside the leucine zipper/basic region.
    Oncogene. 1990 Jun;5(6):929-39 PMID: 2113670
  82. Epidermal growth factor and oncogenes induce transcription of the same cellular mRNA in rat fibroblasts.
    EMBO J. 1985 Jun;4(6):1435-40 PMID: 3875482
  83. Epidermal growth factor stimulation of stromelysin mRNA in rat fibroblasts requires induction of proto-oncogenes c-fos and c-jun and activation of protein kinase C.
    Mol Cell Biol. 1990 Aug;10(8):4284-93 PMID: 2115124
  84. Targeted degradation of c-Fos, but not v-Fos, by a phosphorylation-dependent signal on c-Jun.
    Science. 1992 Dec 18;258(5090):1941-4 PMID: 1470918
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.14.6.4295-4310.1994
Published
1994-06-00
Pages
4295-310
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC358796
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.14.6.4295-4310.1994 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2