Nucleosome core particles of calf thymus, Tetrahymena, and the reconstituted hybrid. Their structure reflects the nature of the histone octamer.
The circular dichroism spectra and the thermal denaturation profiles of the nucleosome core particles isolated by micrococcal nuclease digestion from nuclei of calf thymus and the protozoan Tetrahymena pyriformis were compared with those of the homogeneous and hybrid core particles reconstituted from calf core DNA and either calf or Tetrahymena histone octamer. The core DNA was obtained from the calf core particle, and both the histone octamers were reconstituted from the acid-extracted four core histones of calf thymus or Tetrahymena, whose amino acid sequences show the largest differences hitherto known. The reconstituted homogeneous core particle was identical in both the physical properties with the isolated calf core particle, showing that the correct reconstitution was achieved. The circular dichroism spectra of the calf and Tetrahymena core particles and the hybrid core particle showed no essential differences, indicating that the three core particles have the same overall structure. The derivative thermal-denaturation profiles, however, clearly differed; the calf core particle showed two melting transitions at 60 degrees C and 72 degrees C, while the Tetrahymena and hybrid core particles showed the same three transitions at 48-50 degrees C, 60-61 degrees C, and 72 degrees C. Thus, the thermal denaturation properties of nucleosome core particles do not reflect the nature of DNA, but rather that of the histone octamer bound to the DNA. We conclude that the Tetrahymena histones are more weakly bound to the DNA than the calf thymus histones in the same overall structure of nucleosomes.
Kasai
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K
Hayashi
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Iwai
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D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D002845:Chromatography / Q000379:methods; D002942:Circular Dichroism; D004247:DNA / Q000032:analysis*; D006657:Histones / Q000302:isolation & purification; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D009707:Nucleosomes / Q000032:analysis*; D011489:Protein Denaturation; D013769:Tetrahymena pyriformis / Q000648:ultrastructure*; D013816:Thermodynamics; D013950:Thymus Gland / Q000648:ultrastructure*
D006657:Histones; D009707:Nucleosomes; D004247:DNA