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Physico-chemical studies on the structure of eukaryotic chromatin

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  • It has been shown that about 50 % of the whole chromatin DNA exists in a very compact form in calfthymus chromatin.These compact fragments can be extracted from the whole chromatin as nuclease resistant fragments by limited nuclease digestion. These fragments involve four out of five histones in folding the DNA ( histone F-1 is not present in these fragments). Using tryptic digestion, which is strictly limited with these complexes, it has been shown that DNA in these fragments is in a folded form and is definitely not in the B-form.Physical methods like circular dichroism measurments,sedimentation velocity and sedimentation equilibrium experiments and thermal denaturation studies have been used to study the unfolding effects of tryptic digestion on these fragments. A method has been established for obtaining clean nuclei from chicken erythrocyte in good yield using a non-ionic detergent P-40.It has been shown that chicken erythrocyte chromatin also contains about 50% nuclease resistant fragments,which are insoluble in low-salt solution.The presence of F-1 seems to be the cause of this insolubility.
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