Salt effects on histone subunit interactions as studied by fluorescence spectroscopy.
The salt concentration dependence of the aggregation properties of calf thymus and chicken erythrocyte histones has been investigated by using fluorescence spectroscopy. The isolated H2A/H2B and H3/H4 subunit preparations were labeled with 5-(dimethylamino)naphthalene-1- sulfonyl (dansyl). This long-lived fluorescence probe allows for the observation of rotations due to tumbling of the particle and thus is a probe for changes in the size of macromolecular assemblies. The fluorescence polarization and lifetime were measured as a function of salt concentration for these isolated preparations. Next, each labeled preparation was reconstituted with its unlabeled complement, and the salt concentration dependence of histone core octamer interactions was investigated in the same manner. Salt-induced core particle formation was observed by monitoring the dansyl-labeled dimers for both the calf thymus and chicken erythrocyte preparations. Evidence for subunit dissociation of the isolated H2A-H2B preparations was also found, as well as aggregation of the isolated H3/H4 subunits to at least dimers of tetramers. The calf thymus H3/H4 preparation was in aggregated form under all conditions studied, whereas the chicken erythrocyte H3/H4 only formed aggregates at high protein or salt concentrations. We have found evidence that the dimer can displace the tetramer from the higher order aggregate in order to form core particles. Such competition between the subunit interfaces in the histone system suggests that they may play a regulatory role in histone-DNA interactions.
Royer
C A
CA
Department of Physics, University of Illinois at Urbana-Champaign, Illinois 61801.
Rusch
R M
RM
Scarlata
S F
SF
D000818:Animals; D002417:Cattle; D002645:Chickens; D003619:Dansyl Compounds; D004912:Erythrocytes / Q000032:analysis; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D006657:Histones / Q000097:blood / Q000302:isolation & purification; D011487:Protein Conformation; D012492:Salts; D013050:Spectrometry, Fluorescence; D013950:Thymus Gland / Q000032:analysis
D003619:Dansyl Compounds; D005456:Fluorescent Dyes; D006657:Histones; D012492:Salts