Modification of Chromatin Structure Following Exposure of MOLT4 Cells to the Carcinogenic Chromium(VI)

  • Subhendra N. Mattagajasingh
  • Hara P. Misra

Abstract

DNA-protein complexes-induced by potassium chromate in human leukemic T-lymphocyte MOLT4 cells were isolated by ultracentrifugal sedimentation in the presene of 2% sodium dodecyl sulfate (SDS) and 5 M urea. The complexes were analyzed by two-dimensional SDS-polyacrylamide gel electrophoresis (PAGE). Three acidic proteins of 74, 44 and 42 kD, and a basic protein of 51 kD were primarily complexed to DNA following 25 μM chromate treatment indicating selectivity in chromate-induced DNA-protein complexes. Higher concentrations of chromate cross-linked many other proteins to DNA. A 43 kD protein predominantly localized in the cytoplasmic fraction was found to be cross-linked to DNA upon chromate treatment. Partial N-terminal amino acid sequencing of p43 showed that it could be a human lectin. Treatment of the complexes with DNase I, RNase and EDTA revealed that sedimentation of the proteins was not due to formation of protein aggregates, but due to their association with DNA. The complexes were disrupted, to some extent, by EDTA indicating the involvement of a chelatable form of chromium in the complex. Because chromate-induced DNA-protein complexes are resistant to treatments such as 2% SDS and 5 M urea, but disrupted under gel electrophoretic conditions, it is possible that chromium could be used as a cross-linking agent for identification of other proteins such as transcription factors, that interact with DNA.

Keywords

Sodium Dodecyl Sulfate MOLT4 Cell Potassium Chromate Chelatable Form Nuclear Matrix Protein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Arslan, P., Beltrame, M., and Tomasi, A., 1987, Intracellular chromium reduction, Biochem. Biophys. Acta, 931: 10–15.CrossRefGoogle Scholar
  2. Bedinger, P., Hochstrasser, M., Jongenel, C.V., and Alberts, B.M., 1983, Properties of the T4 bacteriophage DNA replication apparatus: the T4 dda DNA helicase is required to pass a bound RNA polymerase molecule, Cell, 34: 115–123.PubMedCrossRefGoogle Scholar
  3. Bouck, N.P., and Benjamin, B.K., 1989, Loss of cancer suppressors, a driving force in carcinogenesis, Chem. Res. Toxicol, 2: 1–11.Google Scholar
  4. Bouliakas, T., 1986, Protein-protein and protein-DNA interactions in calf thymus nuclear matrix using cross-linking by ultraviolet irradiation, Cell Biol, 64: 474–484.Google Scholar
  5. Briggs, J.A., and Briggs, R.C., 1988, Characterization of chromium effects on a rat liver epithelial cell line and their relevance to in vitro transformation, Cancer Res, 48: 6484–6490.PubMedGoogle Scholar
  6. Chiu, S., Friedman, L., Sokany, N., Xue, L., Oleinick, N., 1986, Nuclear matrix proteins are crosslinked to transcriptionally active gene sequences by ionizing radiation, Radiat. Res, 107: 24–38.Google Scholar
  7. Cohen, M., Latta, D., Coogan, T., and Costa, M., 1990, Mechanisms of Metal carcinogenesis: The reactions of metals with nucleic acids, In: E.C. Foulkes (ed.) The Biological effects of heavy metals, Vol. II, CRC Press, Boca Raton, Fl,.pp 19–76.Google Scholar
  8. De Flora, S., and Wetterhahn, K.E., 1989, Mechanisms of chromium metabolism and genotoxicity, Life. Chem. Rep, 7: 169–277.Google Scholar
  9. Fornace, A.J., Jr., Seres, D.S., Lechner, J.F., and Harris, C.C., 1981, DNA-protein cross-linking by chromium salts, Chem.-Biol. Interact, 36: 345–354.CrossRefGoogle Scholar
  10. Gabius, H., Engelhardt, R., Graupner, G., and Cramer, F., 1986, Lectins in carcinoma cells: Level reduction as possible regulatory event in tumor growth and colonization, In: Lectins: Biology, Biochemistry, Clinical Biochemistry. Bog-Hansen, T.C., and van Driessche, E. (eds) vol. 5, Walter de Gruyter amp; Co., Berlin, pp. 237–242.Google Scholar
  11. Hamilton, J.W., and Wetterhahn, K.E., 1989, Differential effects of chromium(VI) on constitutive and inducible gene expression in chick embryo liver in vivo and correlation with chromium(VI)-induced DNA damage, Mol. Carcinog. 2: 274–286.PubMedCrossRefGoogle Scholar
  12. Kawanishi, S., Inoue, S., Sano, S., 1986, Mechanism of DNA cleavage induced by sodium chromate(VI) in the presence of hydrogen peroxide, J. Biol. Chem. 261: 5952–5989.PubMedGoogle Scholar
  13. Kolb-Bachofen, V., 1986, Mammalian lectins and their function–A Review, In: Bog-Hansen, T.C., and van Driessche, E. (eds), Lectins: Biology, Biochemistry, Clinical Biochemistry (de Gruyter, Berlin ), pp. 197–206.Google Scholar
  14. Koster. A., and Beyersmann, D., 1985, Chromium binding by calf thymus nuclei and effects on chromatin, Toxicol. Environ. Chem. 10: 307–313.Google Scholar
  15. Lesko, S.A., Drocourt, J., Yang, S., 1982, Deoxyribonucleic acid-protein and deoxyribonucleic acid interstrand cross-links induced in isolated chromatin by hydrogen peroxide and ferrous ethylenediaminetetraacetate chelates, Biochemistry 21: 5010–5015.PubMedCrossRefGoogle Scholar
  16. Lis, H., and Nathan, S., 1986(a), Application of lectins, In: The lectins: Properties, function, and applications in biology and medicine, Liener, I.E., Sharon, N., and Goldstein, I.J. (eds) Academic press, Inc., New York, pp. 293–370.Google Scholar
  17. Lis, H., and Nathan, S., 1986(b), Biological properties of lectins, In: The lectins: Properties, function, and applications in biology and medicine, Liener, I.E., Sharon, N., and Goldstein, I.J. (eds) Academic press, Inc., New York, pp. 265–291.Google Scholar
  18. Maniatis, T., Fritsch, E.F., and Sambrook, J. (eds.) 1982, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Pubications, New York, New York.Google Scholar
  19. Mattagajasingh, S.N., and Misra, H.P., 1994, Partial Sequencing of a Protein Cross-linking to DNA upon treatment of Cultured Intact Human Cells (MOLT4) with the Carcinogen Chromium(VI), J. Protein Chem. 13: 449–450.Google Scholar
  20. Mattagajasingh, S.N., and Misra, H.P., 1993, Vitamin E suppresses the potassium chromate induced crosslinking of proteins to DNA in MOLT4 cells, FASEB J. 7: 469.Google Scholar
  21. Miller, C.A., III, and Costa, M., 1989, Analysis of proteins cross-linked to DNA after treatment of cells with formaldehyde, chromate, and cis-diamminedichroloplatinum(II), Mol. Toxicol. 2: 11–26.Google Scholar
  22. O’Farrell, P.Z., Goodman, H.M., and O’Farrell, P.H., 1977, High resolution two dimensional electrophoresis of basic as well as acidic proteins, Cell 12: 1133–1142.PubMedCrossRefGoogle Scholar
  23. Oleinick, N.L., Chiu, S., Ramakrishnan, N., and Xue, L., 1987, The formation, identification, and significance of DNA-protein cross-links in mammalian cells, Br. J. Cancer, (suppl. 8 ) 55: 135–140.Google Scholar
  24. Ono, H., Wada, O., and Ono, T., 1981, Distribution of trace metals in nucleoli of normal and regeneratin rat liver with special reference to the different behavior of nickel and chromium, J. Toxicol. Environ. Health, 8: 947–957.Google Scholar
  25. Sammons, D.W., Adams, L.D., and Nishiziwa, E.E., 1981, Ultracensitive silver based color staining of polypeptides in polyacrylamide gels, Electrophoresis. 2: 135–141.CrossRefGoogle Scholar
  26. Shi, X., and Dalai, N.S., 1989, Chromium(V) and hydrogen radical formation during the glutathione reductase-catalyzed reduction of chromium(VI), Biochem. Biophys. Res. Commun, 163: 627–634.Google Scholar
  27. Standeven, A.M., and Wetterhahn, K., 1991, Is There a Role for Reactive Oxygen Species in the Mechanism of Chromium(VI) Carcinogenesis?, Chem. Res. Toxicol, 4: 616–625.PubMedCrossRefGoogle Scholar
  28. Stein, G.S., and Kleinsmith, L.J., eds., 1979, Chromosal Proteins and Their Role in Regulation of Gene Expression, Academic Press, New York.Google Scholar
  29. Sugiyama, M., Patiemo, S.,R., Catoni, O., and Costa, M., 1986, Characterization of DNA lesions induced by CaCrO4 in synchronous and asynchronous cultured mammalian cells, Mol. Phannacol, 29: 606–613.Google Scholar
  30. Tsapakos, M.J., and Wetterhahn, K.E., 1983, The interaction of chromium with nucleic acids, Chem.-Biol. Interact, 46: 265–277.PubMedCrossRefGoogle Scholar
  31. Wedrychowski, A., Schmidt, W.N., and Hnilica, L.S., 1986, The in vivo crosslinking of proteins and DNA by heavy metals, J. Biol. Chem, 261: 3370–3376PubMedGoogle Scholar
  32. Wedrychowski, A., Ward, W.S., Schmidt, W.N., and Hnilica, L.S., 1985, Chromium-induced cross-linking of nuclear proteins and DNA, J. Biol. Chem, 260: 7150–7155.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Subhendra N. Mattagajasingh
    • 1
  • Hara P. Misra
    • 2
  1. 1.School of MedicineJohn Hopkins UniversityBaltimoreUSA
  2. 2.Department of Biomedical Scicences and Pathobiology Virginia-Maryland Regional College of Veterinary MedicineVirginia Polytechnic Institute and State UniversityBlacksburgUSA

Personalised recommendations