Core nucleosomes by digestion of reconstructed histone-DNA complexes

Nucleic Acids Res. 1979 Apr;6(4):1449-65. doi: 10.1093/nar/6.4.1449.

Abstract

Reconstructed complexes of the inner histones (H2A, H2B, H3, H4) and a variety of DNAs were digested with micrococcal nuclease to yield very homogeneous populations of core nucleosomes (nu 1). Nucleosomes containing Micrococcus luteus DNA (72% G+C); chicken DNA (43% G+C), Clostridium perfringens DNA (29% G+C); or poly(A-dT.poly(dA-dT) have been examined by circular dichroism, thermaldetenaturation, electron microscopy, and DNAse I digestion. Circular dichroism spectra of all particles show a typically suppressed ellipticity at 260--280 nm and a prominent alpha-helix signal at 222 nm. All particles show biphasic melting except nu 1 (dA-dT), which show three prominent melting transitions at ionic strength less than or equal to 1 mM. DNAse I digestion of nu 1 (dA-dT) produces a ladder of DNA fragments fiffering in lengthy by one base residue. nu 1 (dA-dT) contain 146 base pairs of DNA and exhibit an average DNA helix pitch of 10.4-10.5 bases per turn. There appear to be two regions of different DNA pitch wihtin nu 1 (dA-dT). It is suggested that the two regions of DNA pitch might correspond to the two regions of the melting profiles.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Nucleus / analysis
  • Chickens
  • Chromatin / ultrastructure*
  • Circular Dichroism
  • DNA* / blood
  • Deoxyribonucleases
  • Deoxyribonucleoproteins* / blood
  • Erythrocytes / analysis
  • Histones* / blood
  • Micrococcal Nuclease
  • Nucleic Acid Conformation
  • Nucleic Acid Denaturation
  • Nucleoproteins* / blood
  • Poly dA-dT

Substances

  • Chromatin
  • Deoxyribonucleoproteins
  • Histones
  • Nucleoproteins
  • Poly dA-dT
  • DNA
  • Deoxyribonucleases
  • Micrococcal Nuclease