Hybridization of Nuclear Matrix Attached Deoxyribonucleic Acid Fragments

Joseph Basler, Ronald Berezney, Nicholas D. Hastie, Dennis Pietras, Sei Ichi Matsui, Avery A. Sandberg

Research output: Contribution to journalArticle

80 Scopus citations

Abstract

Annealing studies were performed on DNA fragments associated with rat and mouse liver interphase nuclear matrix and the metaphase scaffold of Chinese hamster DON cells. Matrix and scaffold bound DNA fragments, reassociated with an excess of total genomic DNA, displayed kinetics virtually identical with total nuclear DNA probes. Moreover, both the extent and kinetics of these hybridizations were independent of the matrix DNA fragment size (<350- 5000 base pairs) and the method of nuclease digestion used in their preparation (DNase I, micrococcal nuclease or endogenous digestion). The repetitive DNA component of the matrix DNA was examined by reacting discrete sizes of matrix DNA fragments (<350-5000 base pairs) from mouse liver with a library of cloned repetitive sequence DNA fragments which included mouse major satellite sequences. Our results demonstrate that short DNA fragments anchored to the nuclear matrix contain these cloned sequences in similar proportion to total nuclear DNA and, when viewed in light of the annealing results, indicate that matrix DNA is not enriched in either repetitive or unique sequences. Furthermore, the matrix DNA fragments appear to contain the entire sequence complexity of the genome. Finally, we hybridized both matrix and total nuclear DNA fragments with cDNA to total nuclear polyadenylated RNA. The kinetics and extent of hybridization indicate that most, if not all, of the actively transcribed DNA sequences are present in similar concentrations. We conclude that in the overall organization of eukaryotic DNA within the nucleus, the repeating domains or loops which have been demonstrated by a number of investigators are not anchored at specific attachment sequences in interphase cells or during mitosis. These findings are discussed with regard to current concepts of eukaryotic DNA loop organization.

Original languageEnglish (US)
Pages (from-to)6921-6929
Number of pages9
JournalBiochemistry
Volume20
Issue number24
DOIs
StatePublished - Nov 1981
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'Hybridization of Nuclear Matrix Attached Deoxyribonucleic Acid Fragments'. Together they form a unique fingerprint.

  • Cite this

    Basler, J., Berezney, R., Hastie, N. D., Pietras, D., Matsui, S. I., & Sandberg, A. A. (1981). Hybridization of Nuclear Matrix Attached Deoxyribonucleic Acid Fragments. Biochemistry, 20(24), 6921-6929. https://doi.org/10.1021/bi00527a027