Mobile histone tails in nucleosomes. Assignments of mobile segments and investigations of their role in chromatin folding.
The 13C NMR spectrum of isolated nucleosome core particles contains many sharp resonances, including resonances of alpha- and beta-carbons, indicating that certain terminal segments of histones rich in basic residues are highly mobile (Hilliard, R. R., Jr., Smith, R. M., and Rill, R. L. (1986) J. Biol. Chem. 261, 5992-5998). Specific histone termini can be removed sequentially from nucleosome core particles by mild treatment with alpha-chymotrypsin or chymotrypsin plus trypsin (Rosenberg, N. L., Smith. R. M., and Rill, R. L. (1986) J. Biol. Chem. 261, 12375-12383). Comparisons of the 13C NMR spectra of native and several partially proteolyzed core particles indicated that a minimum of residues 1-20 of H3 and 1-11 and 118-128 of H2a are contained in mobile segments of native cores. H4 did not appear to contribute to the resonances from mobile histone segments, but a possible contribution of H2b residues 1-16 could not be ruled out. The 13C NMR spectra of oligonucleosomes containing and lacking lysine-rich histones (H1, H5) were similar to each other and to that of native nucleosome cores both when the oligonucleosomes were in an extended conformation at low ionic strength and when they were in a more compact conformation at higher ionic strength. This similarity suggests that histones H1 and H5 must be largely immobilized upon chromatin binding and that the segments of core histones that are mobile in isolated nucleosome cores are not strongly bound to adjacent linker regions in intact chromatin, and are not immobilized by compaction to the degree achieved in 50 mM phosphate buffer.
Smith
R M
RM
Department of Chemistry, Florida State University, Tallahassee 32306-3006.
Rill
R L
RL
D000818:Animals; D002247:Carbon Isotopes; D002645:Chickens; D002843:Chromatin / Q000378:metabolism* / Q000648:ultrastructure; D004912:Erythrocytes / Q000378:metabolism; D006657:Histones / Q000378:metabolism* / Q000648:ultrastructure; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009707:Nucleosomes / Q000378:metabolism* / Q000648:ultrastructure; D011487:Protein Conformation
D002247:Carbon Isotopes; D002843:Chromatin; D006657:Histones; D009707:Nucleosomes