Effects of shearing on chromatin structure

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Abstract
The effects of mechanical shearing on [calf thymus] chromatin structure were investigated by using thermal denaturation and circular dichroism (CD) spectroscopy. Under ordinary conditions of mechanical shearing used for preparation of soluble chromatin, only minor changes (less than 10%) of chromatin properties were observed with respect to absorption melting curves, CD spectra, CD melting curves and histone transfer from chromatin to exogenous DNA. Such small perturbation of structural properties could be due to the generation of free ends when a large chromatin was cut into smaller fragments and by weakening the binding of histones to DNA near these free ends. In addition to mechanical shearing, sonication was used to shear some samples of chromatin. The effect of sonication on chromatin structure was investigated by the same physical methods used for mechanically sheared chromatin. Sonication only slightly changes the chromatin properties with respect to CD spectra, similar to the results obtained by mechanical shearing, but sonication at high settings has a greater effect on the thermal denaturation property of chromatin in contrast to results from mechanically sheared chromatin.

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