Chromatin fiber nucleosome stability

Figure 3. The stability of the nucleosome is affected by the length and the superhelicity of DNA. (a-b) The chromatin fibers were reconstituted from the purified plasmids and the histone octamers by a salt-dialysis method and observed under AFM. The 3 kb (a) or 106 kb (e) supercoiled circular plasmid was used as a template, (c) Relationship between the plasmid length and the frequency of nucleosome formation in the reconstitution process. The nucleosome frequency is represented as the number of base pairs per nucleosome and plotted against the length of the template DNA in supercoiled (filled circle) and linear (open circle) forms, (d) AFM image of the chromatin fiber reconstituted on the topoisomerase 1-treated plasmid, (e) Chromatin fiber reconstituted with Drosophila embryo extract. The chromatin fiber was reconstituted from plasmid DNA of 10kband the embryo extract of Drosophila, and was observed by AFM...

Mg (but not Na" ") results in a structure that is equivalent to the 30-nm compact fiber in the extent of condensation [49]. Finally, the independent and critical function of core histone N-termini in chromatin condensation was demonstrated by showing that nucleosomal filaments reconstituted from core histones lacking their N-terminal domains are unable to condense into folded structures upon an increase of Mg " ", despite the presence of properly bound histone H5 ([50,51], see also Ref. [52] for the discussion of the special role of H3 and H4 tails). Thus, the presence of HI is not a sine-qua-non condition for salt-induced chromatin folding, which can proceed in Hi s absence and is an intrinsic property of filaments consisting of spaced core particles. A key question is how many of the features of the native 30-nm compact fiber are due to the presence of histone HI From the available data it seems that HI may influence the intrinsic folding pathway of the chromatin filament by stabilizing a single ordered conformation. This property can have much to do with the cooperativity of HI interactions within chromatin but also with the way HI is bound to the nucleosome and with the efifect it exerts on the path of linker DNA. 

In the sections that follow, we are going to describe a few representative examples of how histone variability (histone variants and their post-translational modifications) can affect nucleosome stability and folding. This data supports the notion that while some of this variability may be exclusively used to provide an informational code [121,123,165] it can also have important implications for structural aspects involved in the highly dynamic nature of the chromatin fiber. 

In attempts to reconstitute nucleosomes on SV40 circular DNA, Cap-lan et al. (37) observed that if the histones were ADP-rihosylated prior to reconstitution assembly of nucleosomes was inhibited by as much as 80%. If nucleosomes were first allowed to assemble, then modified by ADP-ribosylation, no eflPect on stability was observed. These data led the authors to speculate that ADP-ribosylation of histones prior to nucleosome assembly might be of physiological importance. Perella and Lea (168, 169) have shown that in rat liver nuclei, polyamines cause an increase in histone Hi ADP-ribosylation and histone HI dimer synthesis, which is accompanied by a decrease in core histone ADP-ribosylation. Data such as these have led to speculation that Hi dimer formation may function in the condensation or stabilization of chromatin fibers (35, 119). The data of Lorimer et al. (124) indicate that dimer synthesis is inversely related to the nuclear activity of the poly(ADP-ribose) glycohydrolase. Thus, these authors (124) inferred that the Hl-Hl-polymer complex formation is of a transient nature. As pointed out by Purnell et al. (171), if this crosslink were to function in the stabilization of chromatin, the modified histone Hi would have... 

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