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Plant and Cell Physiology, 1986, Vol. 27, No. 2 293-302
© 1986

Article

Effect of Calcium on the Structure and Template Activity of Pea Chromatin

Hideaki Matsumoto and Toshio Takyu

Institute for Agricultural and Biological Sciences, Okayama University Chuo 2-20-1, Kurashiki, Okayama 710, Japan

The absorption spectrum of native pea chromatin solubilized under minimal shearing conditions changed with increasing Ca2+ concentration; the ratio of maximum to minimum absorption decreased and the maximum absorption peak shifted to a longer wavelength. The concentration of Ca2+ to cause half complete sedimentation of chromatin was much lower for the solubilized native chromatin (more condensed and larger in size) than for the sonicated chromatin (less condensed and smaller in size). Solubilized native chromatin showed a two-step melting profile in the absence of Ca2+.

In the presence of Ca2+ the two Tms disappeared and a new higher Tm appeared. Template activity of solubilized native chromatin increased 3-fold upon dispersion and fragmentation by sonication. Addition of a small amount of ethylene glycol-bis (ß-aminoethyl ether)-N, N'-tetraacetic acid (EGTA) promoted the template activity of solubilized native chromatin, but not that of sonicated or denatured DNA. The effect of EGTA was reversed by Ca2+. The chromatin reconstituted in the presence of EGTA showed a lower Tm than the chromatin reconstituted in the presence of Ca2+. The relationship between chromatin structure and its template activity is discussed in relation to Ca2+.

(Received August 12, 1985; Accepted December 7, 1985)
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