Diversity of nuclear protein fractions of hamster liver and hepatoma produced by DNaseI.
The structural and functional diversity between active and inactive chromatin is thought to depend, in part, upon differences in DNA-bound protein composition, including changes in the number of sulfhydryl groups. The aim of the present study was to compare protein composition in untreated nuclei, DNaseI-released and resistant nuclear fractions of hamster liver and Kirkman-Robbins hepatoma cells. Electrophoretic analysis of nuclear proteins showed some evident quantitative and qualitative differences between normal and neoplastic cells. The most significant diversities were noticed in DNaseI solubilized fraction of both types of cells. Nuclease attack released a characteristic set of non-histones with mol. wt 37,000, 50,000, 74,000 and 130,000-160,000 from transformed cells, and polypeptides of mol. wt 45,000 and 76,000 from normal cells. Cell-specific distribution within examined nuclear polypeptides was revealed using selective staining of their protein-bound sulfhydryls. Immunoblot analysis demonstrated that a non-histone protein with mol. wt 48,000, overexpressed in rodent tumour cells, was exclusively concentrated in liver DNaseI-sensitive fraction, which amounted only to 8.3% +/- 2.0% of total nuclear DNA. In hepatoma cells, however, this particular polypeptide is distributed between nuclease-sensitive and nuclease-resistant nuclear fractions. Non-histone protein of mol. wt 48,000 appeared to contain free sulfhydryl groups. In summary, these results show molecular specificity of nuclear proteins from normal and tumour cells and differences in their distribution among nuclease-released and nuclease-resistant nuclear fractions. The diversity in molecular characteristics and sulfhydryl group patterns observed among the examined proteins of normal and neoplastic cells may suggest their involvement in some changes in the rearrangement of nuclei during neoplastic transformation.
Kiliańska
Z M
ZM
Department of Cytobiochemistry, University of Lódź, Poland.
Szymczyk
P
P
Krajewska
W M
WM
D000818:Animals; D005591:Chemical Fractionation; D006224:Cricetinae; D004268:DNA-Binding Proteins / Q000378:metabolism*; D003850:Deoxyribonuclease I; D004591:Electrophoresis, Polyacrylamide Gel; D008099:Liver / Q000378:metabolism*; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D008647:Mesocricetus; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000032:analysis* / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000032:analysis
D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D013438:Sulfhydryl Compounds; D003850:Deoxyribonuclease I