Formation and characterization of soluble complexes of histone H1 with supercoiled DNA.
We have analyzed the interaction of rat liver histone H1 with superhelical DNA. Depending on the ratio of H1 to DNA and the concentration of salt, two different types of complexes were found. Above a critical ratio of H1 to DNA, called the aggregation point, large aggregates are formed, which have a cable-like appearance in the electron microscope. Below the aggregation point, individual soluble complexes are formed, which are the subject of this study. With increasing ionic strength, the aggregation point is shifted towards lower ratios of H1 to DNA. In the soluble complexes, H1 appears to bind along superhelically intertwined DNA strands, forming a polymer. Partial digestion of the complexes with protease suggests protection of the N-terminal tail and the globular domain of H1. Similar soluble complexes were observed with various H1 fragments but not with the core histones. In the soluble complexes, similar regions of the H1 molecule are considered to be protected from cleavage by protease, as in chromatin. Therefore, these complexes appear to be a valuable model for the interaction of H1 in chromatin fibers.
D000818:Animals; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000378:metabolism; D002918:Chymotrypsin; D004278:DNA, Superhelical / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D006657:Histones / Q000378:metabolism*; D008099:Liver / Q000378:metabolism; D008854:Microscopy, Electron; D009994:Osmolar Concentration; D051381:Rats; D014357:Trypsin
D002843:Chromatin; D004278:DNA, Superhelical; D006657:Histones; D002918:Chymotrypsin; D014357:Trypsin