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Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA.

Publication ,  Journal Article
Barr, FG; Kastan, MB; Lieberman, MW
Published in: Biochemistry
March 12, 1985

We have compared the distribution of 5-methyldeoxycytidine (m5dC) between staphylococcal nuclease (SN) sensitive and resistant regions of human diploid fibroblast chromatin to the corresponding distribution in purified DNA. After SN digestion of fibroblast nuclei or purified DNA, nuclease-resistant products were separated from sensitive products by perchloric acid or ethanol precipitation; the radioactively labeled nucleosides were then fractionated by high-performance liquid chromatography and quantitated. Our results indicate that m5dC is preferentially associated with SN-resistant regions of both chromatin and purified DNA. The magnitudes of these preferences in fibroblast chromatin and DNA are similar; we find that the enrichment of m5dC content in SN-resistant fractions of nuclei and DNA relative to the corresponding sensitive fractions is approximately 2-3-fold. Therefore, highly methylated regions of DNA have an intrinsic resistance to digestion by SN that is of sufficient magnitude to explain the high degree of nuclease resistance of chromatin containing highly methylated DNA.

Duke Scholars

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 12, 1985

Volume

24

Issue

6

Start / End Page

1424 / 1428

Location

United States

Related Subject Headings

  • Micrococcal Nuclease
  • Humans
  • Deoxycytidine
  • DNA
  • Chromatography, High Pressure Liquid
  • Chromatin
  • Cell Nucleus
  • Biochemistry & Molecular Biology
  • 3404 Medicinal and biomolecular chemistry
  • 3205 Medical biochemistry and metabolomics
 

Citation

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Barr, F. G., Kastan, M. B., & Lieberman, M. W. (1985). Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA. Biochemistry, 24(6), 1424–1428. https://doi.org/10.1021/bi00327a021
Barr, F. G., M. B. Kastan, and M. W. Lieberman. “Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA.Biochemistry 24, no. 6 (March 12, 1985): 1424–28. https://doi.org/10.1021/bi00327a021.
Barr FG, Kastan MB, Lieberman MW. Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA. Biochemistry. 1985 Mar 12;24(6):1424–8.
Barr, F. G., et al. “Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA.Biochemistry, vol. 24, no. 6, Mar. 1985, pp. 1424–28. Pubmed, doi:10.1021/bi00327a021.
Barr FG, Kastan MB, Lieberman MW. Distribution of 5-methyldeoxycytidine in products of staphylococcal nuclease digestion of nuclei and purified DNA. Biochemistry. 1985 Mar 12;24(6):1424–1428.
Journal cover image

Published In

Biochemistry

DOI

ISSN

0006-2960

Publication Date

March 12, 1985

Volume

24

Issue

6

Start / End Page

1424 / 1428

Location

United States

Related Subject Headings

  • Micrococcal Nuclease
  • Humans
  • Deoxycytidine
  • DNA
  • Chromatography, High Pressure Liquid
  • Chromatin
  • Cell Nucleus
  • Biochemistry & Molecular Biology
  • 3404 Medicinal and biomolecular chemistry
  • 3205 Medical biochemistry and metabolomics