Restriction enzyme analysis of DNA methylation in "condensed" chromatin of Ha-ras-transformed NIH 3T3 cells.

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  • Author(s): Mello ML;Mello ML; Chambers AF; Vidal BC; Planding W; Schenck U
  • Source:
    Analytical cellular pathology : the journal of the European Society for Analytical Cellular Pathology [Anal Cell Pathol] 2000; Vol. 20 (4), pp. 163-71.
  • Publication Type:
    Comparative Study; Journal Article; Research Support, Non-U.S. Gov't
  • Language:
    English
  • Additional Information
    • Source:
      Publisher: IOS Press Country of Publication: Netherlands NLM ID: 8911016 Publication Model: Print Cited Medium: Print ISSN: 0921-8912 (Print) Linking ISSN: 09218912 NLM ISO Abbreviation: Anal Cell Pathol Subsets: MEDLINE
    • Publication Information:
      Publication: Amsterdam : IOS Press
      Original Publication: Amsterdam : Elsevier Science Publishers, c1989-c2003.
    • Subject Terms:
      DNA Methylation* ; Genes, ras* ; Rosaniline Dyes*; Chromatin/*chemistry ; DNA/*metabolism; 3T3 Cells ; Animals ; Cell Line, Transformed ; Chromatin/genetics ; Coloring Agents ; DNA-Cytosine Methylases ; Deoxyribonuclease HpaII ; Image Processing, Computer-Assisted ; Mice ; Microscopy, Video
    • Abstract:
      Increased amounts of chromatin condensation (i.e., localized areas of high DNA density, or chromatin higher order packing state) have been described in NIH 3T3 cells transformed with the Ha-ras oncogene. The structural basis for this oncogene-mediated alteration in nuclear organization is unknown. Since DNA methylation is likely to be involved in regulating the nucleosomal level of DNA packaging, we studied the role of DNA methylation in higher-order chromatin organization induced by Ha-ras. CpG-methylated DNA content was estimated in "condensed" chromatin of Ha-ras-transformed NIH 3T3 cell lines which differ in ras expression and ras-induced metastatic ability but present approximately the same values of "condensed" chromatin areas. The question posed was that if DNA methylation were involved with the chromatin higher-order organization induced by Ha-ras in these cell lines, the methylated DNA density in the "condensed" chromatin would also be the same. The DNA evaluation was performed by video image analysis in Feulgen-stained cells previously subjected to treatment with Msp I and Hpa II restriction enzymes, which distinguish between methylated and non-methylated DNA. The amount of methylated CpG sequences not digested by Hpa II in "condensed" chromatin regions was found to vary in the studied ras-transformed cell lines. DNA CpG methylation status is thus suggested not to be involved with the higher order chromatin condensation induced by ras transformation in the mentioned NIH 3T3 cell lines.
    • Accession Number:
      0 (Chromatin)
      0 (Coloring Agents)
      0 (Feulgen stain)
      0 (Rosaniline Dyes)
      9007-49-2 (DNA)
      EC 2.1.1.- (DNA modification methylase SssI)
      EC 2.1.1.- (DNA-Cytosine Methylases)
      EC 3.1.21.- (Deoxyribonuclease HpaII)
    • Publication Date:
      Date Created: 20010224 Date Completed: 20010322 Latest Revision: 20191104
    • Publication Date:
      20221213
    • Accession Number:
      PMC4618154
    • Accession Number:
      10.1155/2000/138230
    • Accession Number:
      11205319