c-Ha-rasVal 12 oncogene-transformed NIH-3T3 fibroblasts display more decondensed nucleosomal organization than normal fibroblasts.
We have compared the nucleosomal organization of c-Ha-rasVal 12 oncogene-transformed NIH-3T3 fibroblasts with that of normal fibroblasts by using micrococcal nuclease (MNase) as a probe for the chromatin structure. The bulk chromatin from asynchronously and exponentially growing ras-transformed cells was much more sensitive to MNase digestion than chromatin from the normal cells. Southern hybridization analyses of the MNase digests with probes specific for the ornithine decarboxylase (odc) and c-myc genes showed that the coding and/or 3' end regions of these growth-inducible genes carry a nucleosomal organization both in ras-transformed and normal cells. Studies with cells synchronized by serum starvation showed that in both cell lines the nucleosomal organization of chromatin is relatively condensed at the quiescent state, becomes highly decondensed during the late G1 phase of the cell cycle, and starts again to condense during the S phase. However, in ras-transformed cells the decondensation state stayed much longer than in normal cells. Moreover, irrespective of the phase of the cell cycle the bulk chromatin as well as that of the odc and c-myc genes was more sensitive to MNase digestion in the ras-transformed cell than in the normal fibroblast. Decondensation of the chromatin was also observed in the normal c-Ha-ras protooncogene-transfected cells, but to a lesser extent than in the mutant ras-transformed cells. Whether the increased degree of chromatin decondensation plays a regulatory role in the increased expression of many growth-related genes in the ras-transformed cells remains an interesting object of further study.
Department of Pathology, University of Helsinki.
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Laitinen
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L
Sistonen
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K
Alitalo
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E
Hölttä
D000818:Animals; D002453:Cell Cycle; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D004247:DNA / Q000302:isolation & purification; D004261:DNA Replication; D005347:Fibroblasts / Q000166:cytology / Q000648:ultrastructure; D005434:Flow Cytometry; D011905:Genes, ras; D051379:Mice; D009707:Nucleosomes / Q000648:ultrastructure*; D009955:Ornithine Decarboxylase / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism; D014162:Transfection; D014633:Valine
D009707:Nucleosomes; D004247:DNA; D009955:Ornithine Decarboxylase; D014633:Valine; D013936:Thymidine