| Abstrakt: |
A refined high-resolution map for the linear arrangement of histones along DNA in the nucleosomal core particles has been determined by DNA-protein crosslinking. Histones are aligned on one strand of the 145 bp core DNA in the following order: (5') H2B25,35--H455,65--H375,85,, 95/H488--H2B105, 115--H2A118--H3135, 145/H2A145 (3') (the subscripts indicate the approximate distance in nucleotides of the main histone binding sites from the 5'-end of the core DNA). This suggests a symmetrical and rather autonomous arrangement of the histone tetramer (H3, H4)2 and two dimers (H2A, H2B) on the double-stranded core DNA: H2A/H3--(H2A, H2B)--(H3, H4)2--(H2B, H2A)--H3/H2A. The arrangement of histones on DNA was found to be very similar for the cores isolated from the repressed nuclei of sea urchin sperm and chicken erythrocytes and from the active in transcription and replication Drosophila embryo and yeast nuclei. This indicates that the core nucleosome structure is highly conserved through evolution and that the overall inactivation of chromatin does not affect the primary organization of the cores. A new binding site H2B58 was found for a sea urchin spermal variant of H2B which contains an additional basic segment within the N-terminal part of the molecule. The core isolation procedure was shown to introduce changes into the core structure which are reflected in the appearance of a new binding site H2A75. |
