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The anti-cancer agent distamycin A displaces essential transcription factors and selectively inhibits myogenic differentiation

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Abstract

The anticancer drug, distamycin A, alters DNA conformation by binding to A/T-rich domains. We propose that binding of the drug to DNA alters transcription factor interactions and that this may alter genetic regulation. We have analyzed the effects of distamycin A upon expression of the muscle-specific cardiac and skeletal α-actin genes which have A/T-rich regulatory elements in their promoters. Distamycin A specifically inhibited endogenous muscle genes in the myogenic C2 cell line and effectively eliminated the myogenic program. Conversely, when 10T1/2C18 derived pleuripotential TA1 cells were induced to differentiate in the presence of distamycin A, adipocyte differentiation was enhanced whereas the numbers of cells committing to the myogenic program decreased dramatically. Using the mobility shift assay, distamycin A selectively inhibited binding of two important transcription factors, SRF and MEF2, to their respective A/T-rich elements. The binding of factors Sp1 and MyoD wer ffected. The inhibition of factor binding correlated with a repression of muscle-specific promoter activity as assayed by transient transfection assays. Co-expression of the myoD gene, driven by a distamycin A-insensitive promoter, failed to relieve the inhibition of these muscle-specific promoters by distamycin A. Additionally, SRF and MEF2 dependent promoters were selectively down regulated by distamycin A. These results suggest that distamycin A may inhibit muscle-specific gene expression by selectively interfering with transcription factor interactions and demonstrate the importance of these A/T-rich elements in regulating differentiation of this specific cell type.

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References

  1. Hahn F: Mechanism of action of antimicrobial and antitumor agents. In: J Corcoran, F Hahn (eds). Antibiotics III. Springer-Vaerlag: New York, 1975, p. 75

    Google Scholar 

  2. Kopka M, Yoon C, Goodsell D, Pjura P, Dickerson R: The molecular origin of DNA-drug specificity in netropsin and distamycin. Proc Natl Acad Sci USA 82: 1376–1380, 1985

    PubMed  Google Scholar 

  3. Portugal J, Waring M: Comparison of binding sites in DNA for berenil, netropsin and distamycin: a footprinting study. Eur J Biochem 167: 281–289, 1987

    PubMed  Google Scholar 

  4. Bruzik J, Auble D, deHaseth P: Specific activation of transcription initiation by the sequence specific DNA binding agents distamycin A and Netropsin. Biochemistry 26: 950–956, 1987

    PubMed  Google Scholar 

  5. Straney D, Crothers D: Effect of drug-DNA interactions upon transcription initiation at the lac promoter. Biochemistry 26: 1987–1995, 1987

    PubMed  Google Scholar 

  6. Lown J, Krowicki K, Balzarini J, DeClercq E: Structure-activity relationship of novel oligopeptide anti-viral anti-tumor agents related to Netropsin and Distamycin. J Med Chem 29: 1210–1214, 1986

    PubMed  Google Scholar 

  7. Low C, Drew H, Warring M: Echinomycin and distamycin induce rotation of nucleosome core DNA. Nucleic Acids Res 14: 6785–6801, 1986

    PubMed  Google Scholar 

  8. Broggini M, Ponti M, Ottolenghi S, D'Incalci M, Mongelli N, Mantovani R: Distamycins inhibit the binding of OTF-1 and NFE-1 transfactors to their conserved DNA elements. Nuc Acids Res 17: 1051–1055, 1989

    Google Scholar 

  9. Davis R, Weintraub H, Lassar A: Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell 51: 987–1000, 1987

    PubMed  Google Scholar 

  10. Wright W, Sassoon D, Lin V: Myogenin, a factor regulating myogenesis, has a domain homologous to MyoD. Cell 56: 607–617, 1989

    PubMed  Google Scholar 

  11. Braun T, Buschhausen-Denker G, Bober E, Tannich E, Arnold H: A novel human muscle factor related to but distinct from MyoD1 in-duces myogenic conversion in 10T1/2 fibroblasts. EMBO J 8: 701–709, 1989

    PubMed  Google Scholar 

  12. Braun T, Bober E, Winter B, Rosenthal N, Arnold H: MYF-6, a new member of the human gene family of myogenic determination factors: Evidence for a gene cluster of chromosome 12. EMBO J 9: 821–831, 1990

    PubMed  Google Scholar 

  13. Miner J, Wold B: Herculin, a fourth member of the MyoD family of myogenic regulatory genes. Proc Natl Acad Sci 87: 1089–1093, 1990

    PubMed  Google Scholar 

  14. Rhodes S, Konieczny S: Identification of MRF-4: a new member of the muscle regulatory gene family. Genes Dev 3: 2050–2061, 1989

    PubMed  Google Scholar 

  15. Gossett L, Kelvin D, Sternberg E, Olson E: A new myocyte-specific enhancer binding factor that recognizes a conserved element associated with multiple muscle-specific genes. Mol Cell Biol 9: 5022–5033, 1989

    PubMed  Google Scholar 

  16. Martin J, Schwarz J, Olson E: Myocyte enhancer factor (MEF) 2C: A tissue-restricted member of the MEF-2 family of transcription factors. Proc Natl Acad Sci USA 90: 5282–5286, 1993

    PubMed  Google Scholar 

  17. Breitbart R, Liang C, Smoot L, Laheru D, Mahdavi V, Nadal-Ginard B: A fourth human MEF2 transcription factor, hMEF2D, is an early marker of the myogenic lineage. Development 118: 1095–1106, 1993

    PubMed  Google Scholar 

  18. Andres V, Fisher S, Wearsch P, Walsh K: Regulation of Gax homeobox gene transcription by a combination of positive factors including myo-cytespecific enhancer factor 2. Mol Cell Biol 8: 4272–4281, 1995

    Google Scholar 

  19. Yu Y.-T, Brietbart S, Lee Y, Mahdavi V, Nadal-Ginard B: Human myocyte-specific enhancer factor 2 comprises a group of tissue-restricted MADS box transcription factors. Genes and Dev. 6: 1783–1798, 1992

    PubMed  Google Scholar 

  20. Nurrish S, Treisman R: DNA binding specificity determinants in MADS-box transcription factors. Mol Cell Biol 8: 4076–4085, 1995

    Google Scholar 

  21. Edmondson D, Cheng T.-C, Cserjesi P, Chakraborty T, Olson E: Analysis of the myogenin promoter reveals an indirect pathway for positive auto-regulation mediated by the muscle-specific enhancer factor MEF-2. Cell. 12: 3665–3677, 1992

    Google Scholar 

  22. Molkentin J, Black B, Martin J, Olson E: Cooperative activation of muscle gene expression by MEF2 and myogenic bHLH proteins. Cell 83: 1125–1136, 1995

    PubMed  Google Scholar 

  23. Gustafson T, Kedes L: Identification of multiple proteins which interact with functional regions of the human cardiac a-actin promoter. Mol Cell Biol 9: 3269–3283, 1989

    PubMed  Google Scholar 

  24. Minty A, Kedes L: Upstream regions of the human cardiac actin gene that modulate its transcription in muscle cells: presence of an evolutionary conserved repeated motif. Mol Cell Biol 6: 2125–2136, 1986

    PubMed  Google Scholar 

  25. Muscat G, Perry S, Prentice H, Kedes L: The human skeletal a-actin gene is regulated by a muscle-specific enhancer that binds three nuclear factors. Gene Expr 2: 111–126, 1992

    PubMed  Google Scholar 

  26. Taylor A, Erba H, Muscat G, Kedes L: Nucleotide sequence and expression of the human skeletal α-actin gene: evolution of functional regulatory domains. Genomics 3: 323–336, 1988

    PubMed  Google Scholar 

  27. Boxer L, Prywes R, Roeder R, Kedes L: The Sarcomeric actin CArG-binding factor is indistinguishable from the c-fos serum response factor. Mol Cell Biol 9: 515–522, 1989

    PubMed  Google Scholar 

  28. Sartorelli V, Webster K, Kedes L: Muscle-specific expression of the cardiac alpha-actin gene requires MyoD1, CArG-box binding factor, and Sp1. Genes and Dev 4: 1811–1822, 1990

    PubMed  Google Scholar 

  29. Gustalson T, Taylor A, Kedes L: DNA bending is induced by a transcription factor which interacts with the human c-fos and α-actin promoter. Proc Natl Acad Sci USA 86: 2162–2166, 1989

    PubMed  Google Scholar 

  30. Yaffe D, Saxel O: Serial passaging and differentiation of myogenic cells isolated from dystrophic mouse muscle. Nature 270: 725–727, 1977

    PubMed  Google Scholar 

  31. Chapman A, Knight D, Dieckmann B, Ringold G: Analysis of gene. expression during differentiation of adipogenic cells in culture and hormonal control of the developmental program. J Biol Chem 259: 15548–15555, 1984

    PubMed  Google Scholar 

  32. Miller S, Ito H, Blau H, Torti F: Tumor necrosis factor inhibits human myogenesis in vitro. Mol Cell Biol 8: 2295–2301, 1988

    PubMed  Google Scholar 

  33. Graham F, Van der Eb A: A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52: 456–457, 1973

    PubMed  Google Scholar 

  34. Gorman C, Moffat L, Howard B: Recombinant genomes which express chloramphenicol acetyl-transferase in mammalian cell. Mol Cell Biol 2: 1044–1051, 1982

    PubMed  Google Scholar 

  35. Dignam J, Lebovitz R, Roeder R: Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nuc Acids Res 11: 1475–1489, 1983

    Google Scholar 

  36. Bradford M: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72: 248–254, 1976

    PubMed  Google Scholar 

  37. Gunning P, Hardeman E, Wade R, Ponte P, Bains W, Blau H, Kedes L: Differential patterns of transcript accumulation during human myogenesis. Mol Cell Biol 7: 4100–4114, 1987

    PubMed  Google Scholar 

  38. Ponte P, Gunning P, Blau H, Kedes L: Human actin genes are single copy for α-skeletal and α-cardiac actin but multicopy for β- and γ -cytoskeletal genes: 3' untranslated regions are isotype specific but are conserved in evolution. Mol Cell Biol 3: 1783–1791, 1983

    PubMed  Google Scholar 

  39. Webster K: Regulation of glycolytic enzyme RNA transcription rates by oxygen availability in skeletal muscle cells. Mol Cell Biochem 77: 19–28, 1987

    PubMed  Google Scholar 

  40. Wells D, Hoffman D, Kedes L: Unusual structure, evolutionary conservation of non coding sequences and numerous pseudogenes characterize the human H3.3 histone multigene family. Nucleic Acids Res 15: 2871–2889, 1987

    PubMed  Google Scholar 

  41. Webster K, Muscat G, Kedes L: Adenovirus E1A products suppress myogenic differentiation and inhibit transcription from muscle-specific promoters. Nature 332: 553–556, 1988

    PubMed  Google Scholar 

  42. Fisch T, Prywes R, Roeder R: c-fos Sequences necessary for basal expression and induction by epidermal growth factor, 12-O-tetradecanoyl phorbol-13-acetate, and the calcium ionophore. Mol Cell Biol 7: 3490–3502, 1987

    PubMed  Google Scholar 

  43. Kong Y, Johnson S, Taparowsky E, Konieczny S: Ras p21Va1 inhibits myogenesis without altering the DNA binding or transcriptional activities of the myogenic basic helix loop-helix factors. Mol Cell Biol 10: 5205–5213, 1995

    Google Scholar 

  44. Gunning P, Leavitt J, Muscat G, Ng S-Y, Kedes L: A human betaactin expression vector directs high-level accumulation of anti-sense transcripts. Proc Natl Acad Sci USA 84: 4831–4835, 1987

    PubMed  Google Scholar 

  45. Childs G, Maxson R, Kedes L: Histone gene expression during sea urchin embryogenesis: isolation and characterization of early and late messenger RNAs of Strongylocentrotus purpuratus by gene specific hybridization and template activity. Dev Biol 73: 153–173, 1979

    PubMed  Google Scholar 

  46. Maniatis T, Fritch E, Sambrook J: In: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NJ, 1982

    Google Scholar 

  47. Ito H, Miller S, Akinoto H, Torti S, Taylor A, Billingham M, Torti F: Quantitative evaluation of mRNA levels by the polymerase chain reaction in small cardiac tissue samples. J Mol and Cell Cardiology 23: 1117–1125, 1991

    Google Scholar 

  48. Fried M, Crothers D: Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nuc Acids Res 9: 6505–6525, 1981

    Google Scholar 

  49. Miwa T, Kedes L: Duplicated CArG box domains have positive and mutually dependent regulatory roles in expression of the human α-cardiac actin gene. Mol Cell Biol 8: 2803–2813, 1987

    Google Scholar 

  50. Buckingham M: Actin and myosin multigene families: their expression during the formation of skeletal muscle. Essays in Biochem 20: 77–109, 1985

    Google Scholar 

  51. Emerson CP: Myogenesis and developmental control genes. Curr Opin Cell Biol 2: 1065–1075, 1990

    PubMed  Google Scholar 

  52. Carmon Y, Czosnek H, Nudel U, Shani M, Yaffe D: DNAase I sensitivity of genes expressed during myogenesis. Nuc Acids Res 10: 3085–3098, 1982

    Google Scholar 

  53. Yisraeli J, Adelstein R, Melloul D, Nudel U, Yaffe D, Cedar H: Muscle-specific activation of a methylated chimeric actin gene. Cell 46: 409–416, 1986

    PubMed  Google Scholar 

  54. Maniatis T, Goodbourn S, Fischer J: Regulation of inducible and tissue-specific gene expression. Science 236: 1237–1245, 1987

    PubMed  Google Scholar 

  55. Weintraub H: Assembly and propagation of repressed and derepressed chromosomal states. Cell 42: 705–711, 1985

    PubMed  Google Scholar 

  56. Wilder E, Linzer D: Participation of multiple factors, including proliferin, in the inhibition of myogenic differentiation. Mol Cell Biol 9: 430–441, 1989

    PubMed  Google Scholar 

  57. Blau H, Epstein C: Manipulation of myogenesis in vitro: reversible inhibition by DMSO. Cell 17: 95–108, 1979

    PubMed  Google Scholar 

  58. Li L, Zhou J, James G, Heller-Harrison R, Czech M, Olson E: FGF inactivates myogenic helix-loop-helix proteins through phosporylation of a conserved protein kinase C site in their DNA-binding domains. Cell 71: 1181–1194, 1992

    PubMed  Google Scholar 

  59. Massague J, Cheifetz S, Endo T, Nadal-Ginard B: Type b transforming growth factor is an inhibitor of myogenic differentiation. Proc Natl Acad Sci USA 83: 8206–8210, 1986

    PubMed  Google Scholar 

  60. Olson E: Interplay between proliferation and differentiation within the myogenic lineage. Dev Biol 154: 261–272, 1992

    PubMed  Google Scholar 

  61. Alema S, Tato F: Oncogenes and muscle differentiation: multiple mechanisms of interference. Cancer Biol 5: 147–156, 1994

    Google Scholar 

  62. Bengal E, Ransone L, Scharfmann R, Dwarki V, Tapscott S, Weintraub H, Verma I: Functional antagonism between c-Jun and MyoD proteins: a direct physical association. Cell 68: 507–519, 1992

    PubMed  Google Scholar 

  63. Falcone G, Tato F, Alema S: Distinctive effects of the viral oncogenes myc, erb, fps, and src of the differentiation program of quail myogenic cells. Proc Natl Acad Sci USA 82: 426–430, 1985

    PubMed  Google Scholar 

  64. Olson E, Spizz G, Tainsky M: The oncogenic forms of N-ras or H-ras prevent skeletal myoblast differentiation. Mol Cell Biol 7: 2104–2111, 1987

    PubMed  Google Scholar 

  65. Weintraub H: The MyoD family and myogenesis: redundancy, networks, and thresholds. Cell 75: 1241–1244, 1993

    PubMed  Google Scholar 

  66. Hogan M, Dattagupta N, Crothers D: Transmission of allosteric effects in DNA.Nature 279: 521–524, 1979

    Google Scholar 

  67. Cserjesi P, Lilly B, Bryson L, Wang Y, Sassoon D, Olson E: MHox: a mesodermally restricted homeodomain protein that binds an essential site in the muscle creatine kinase enhancer. Development 115: 1087–1101, 1992

    PubMed  Google Scholar 

  68. Duprey P, Lesens C: Control of skeletal muscle-specific transcription: involvement of paired homeodomain and MADS domain transcription factors. Int J Dev Biol 3X: 591–604, 1994

    Google Scholar 

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Authors and Affiliations

  1. Department of Biological Sciences, Wichita State University, Wichita, KS, 67208, USA

    Alan Taylor

  2. Department of Cell and Molecular Biology, SRI International, Menlo Park, CA, 94025, USA

    Keith A. Webster

  3. Department of Physiology, University of Maryland, School of Medicine, Baltimore, MD, 21201, USA

    Thomas A. Gustafson

  4. Institute for Genetic Medicine, University of Southern California School of Medicine, Los Angeles, CA, 90033, USA

    Larry Kedes

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  1. Alan Taylor
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  2. Keith A. Webster
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  4. Larry Kedes
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Taylor, A., Webster, K.A., Gustafson, T.A. et al. The anti-cancer agent distamycin A displaces essential transcription factors and selectively inhibits myogenic differentiation. Mol Cell Biochem 169, 61–72 (1997). https://doi.org/10.1023/A:1006898812618

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