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

  • Subhendra N. Mattagajasingh
  • Hara P. Misra


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.


Sodium Dodecyl Sulfate MOLT4 Cell Potassium Chromate Chelatable Form Nuclear Matrix Protein 
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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

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