Studies on synthetic chromatins containing poly(dA-dT) X poly(dA-dT) and poly(dG-dC) X poly(dG-dC).

Academic Article

Abstract

  • Core histones (H2A,H2B,H3,H4)2, were reconstituted with the synthetic polynucleotides poly(dA-dT) X poly(dA-dT) and poly(dG-dC) X poly(dG-dC) to yield synthetic chromatins containing 200 basepairs per octamer. These synthetic chromatins displayed a 36% decrease in the circular dichroism (CD) peak ellipticity from the value of the polynucleotide free in solution; the poly(dA-dT) X poly(dA-dT)/chromatin showed an increase in the complexity of the thermal denaturation profile compared to that of the polynucleotide. Both the temperature of maximum dh/dT for each transition (Tm) and the relative amount of poly(dA-dT) X poly(dA-dT) in the synthetic chromatin melting in each of the four thermal transitions is a function of the ionic strength over the 0-5 mM sodium phosphate range (0.25 mM EDTA, pH 7.0); a shift of material toward higher melting transitions was observed with increasing ionic strength. The CD peak ellipticity value for both synthetic chromatins was ionic strength-independent over the 0-5 mM sodium phosphate range. These results are in contrast to those observed with H1/H5 stripped chicken erythrocyte chromatin (Fulmer, A. and Fasman, G.D. (1979) Biopolymers 18, 2875-2891), where an ionic strength dependence was found. Differences in the CD spectra between poly(dA-dT) X poly(dA-dT)/chromatin, poly(dG-dC) X poly(dG-dC)/chromatin and H1/H5 stripped chicken erythrocyte chromatin suggest subtle differences in assembly. Finally, the temperature dependence of the CD spectra of poly(dA-dT) X poly(dA-dT)-containing synthetic chromatin, which is similar to that for the polynucleotide, suggests the core histone bound polynucleotide has a large degree of conformational flexibility allowing it to undergo the premelt transition.
  • Keywords

  • Animals, Chickens, Chromatin, Circular Dichroism, Histones, Models, Genetic, Nucleic Acid Conformation, Poly dA-dT, Polydeoxyribonucleotides, Protein Conformation, Spectrophotometry, Ultraviolet
  • Digital Object Identifier (doi)

    Author List

  • Prevelige PE; Fasman GD
  • Start Page

  • 85
  • End Page

  • 96
  • Volume

  • 739
  • Issue

  • 1