Identification of attachment proteins for DNA in chinese hamster ovary cells

A. E. Cress, K. M. Kurath

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

Protein candidates for the attachment of DNA within eukaryotic cell nuclei were identified by isolating nuclear matrix proteins and determining which of those proteins co-sedimented with DNA within a 5.7 M CsCl gradient. The presence of attached nucleic acid was detected after the proteins were subjected to the denaturing conditions of isoelectric focusing/sodium dodecyl sulfate two-dimensional polyacrylamide gel electrophoresis. The attached nucleic acid was detected with silver staining, ethidium bromide, and Amido Black binding. The nucleic acid was identified as DNA based on its ability to be labeled in vitro by terminal deoxynucleotidyltransferase and DNA polymerase I (Klenow). Three proteins were identified as containing attached DNA, one of which was vimentin. The proteins had apparent M(r) and pI values of 70,000, 4.3; 70,000, 5.3; and 57,000, 4.8, respectively. We propose that these proteins are within a class of nuclear proteins containing firmly attached DNA and have referred to them as DNA attachment proteins.

Original languageEnglish (US)
Pages (from-to)19678-19683
Number of pages6
JournalJournal of Biological Chemistry
Volume263
Issue number36
StatePublished - Dec 1 1988

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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