Histone crosslinking patterns indicate dynamic binding of histone H1 in chromatin.

Crosslinking of histone H1 molecules to each other and to the core histones with bifunctional reagents in mouse liver nuclei and chromatin was compared with that under the conditions of random 'contacts' between these molecules. The patterns of crosslinking of the H1 subfractions (H1A, H1B, and H10) to each other in nuclei, chromatin and in solution at different ionic strengths due to random collisions were essentially the same. Moreover, the contacts between the H1 molecules were qualitatively the same in nuclei, chromatin and in solution also at the level of the chymotryptic halves of the H1 molecules. The contacts between the H1 molecules and the core histones in nuclei were similar to those obtained in chromatin at 70 mM NaCl, when H1 molecules readily migrate, and at 0.6 M NaCl, when H1 molecules are dissociated from chromatin. We conclude that spatial arrangement of H1 subfractions and mutual orientation of H1 molecules in isolated nuclei are random-like at least in terms of cross-linking. The static and dynamic models of histone H1 binding to chromatin compatible with the known data are considered. Although unequivocal verification of the models is not possible at present, the dynamic models do correspond better to recent data on the location of the histone H1 in nuclei and chromatin.

[Localization of histone H1 in chromatin. Cross-linking of the N- and C-terminal halves of the molecule with bifunctional reagents]. / Raspolozhenie gistona H1 v khromatine. Sshivanie bifunktsional'nym reagentom N- i C-kontsevykh polovin molekuly.

Mutual arrangement of histone H1 molecules was studied in calf thymus nuclei, extended chromatin and chromatin, isolated and kept in 8 M urea. Histone H1 dimers crosslinked with methyl 4-mercaptobutyrimidate were digested with chymotrypsin and crosslinked fragments obtained were analysed by diagonal gel electrophoresis. In all chromatins tested the N- and C-terminal parts of the H1 molecules were crosslinked in all possible combinations, i.e. C-C, C-N and N-N. These and related data obtained earlier indicate, that the proximity of histone H1 molecules in chromatin is determined by the structure of nucleosomal chain itself and not by chromatin superstructure. The results also suggest that the H1A and H1B subfractions of histone H1 are interspersed in extended nucleosomal chains.

[Localization of histone H1 in chromatin. Cross-linking of central globular regions of H1 molecules with a bifunctional reagent]. / Raspolozhenie gistona H1 v khromatine. Poperechnoe sshivanie tsentral'nykh globuliarnykh chastei molekul H1 bifunktsional'nym reagentom.

Mutual arrangement of histone H1 molecules and central globular parts of H1 was studied by crosslinking with a reversible bifunctional reagent. The yields of histone H1 dimers and dimers of it's globular fragment in nuclei and isolated chromatin were similar. In the presence of 8 M urea the yield of the H1 dimers was approximately threefold decreased, dimers of globular fragment being practically absent. The data suggest that the proximity of H1 molecules in nuclei is stipulated by a structure of a nucleosomal chain itself and not by chromatin superstructure. The results are in accord with the "head" to "head" histone H1 orientation within the nucleosomal chain and do not support participation of the central globular region of H1 molecule in chromatin condensation. A model of H1 arrangement in extended nucleosomal chain is proposed.

Mutual arrangement of histone H1 molecules in extended chromatin. Chymotryptic digestion of cross-linked H1 histone dimers.

Mutual arrangement of histone H1 molecules in chromatin extended in low salt-EDTA buffer and additionally in the presence of urea was studied by means of reversible cross-linking combined with chymotryptic digestion. In the chromatins tested, the chymotryptic halves of H1 were cross-linked in all possible combinations; i.e., C-C, C-N and N-N. The results imply that the mutual arrangement of H1 histones is determined by the structure of extended nucleosomal chain, rather than chromatin superstructure.

Chemical crosslinking of histone H1o to histone neighbours in nuclei and chromatin.

Crosslinking of histones in mouse liver nuclei and extended chromatin with a bifunctional reagent leads to the formation of H1-H1o heterodimers as well as H1o-H1o homodimers. H1o can be also crosslinked to the core histones. Thus, the location of histone H1o within the basic repeating chromatin structure seems to be analogous to that of H1 histone.