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Nucleic Acids Research, 1978, Vol. 5, No. 10 3881-3898
© 1978

Articles

Nucleosome conformation: pH and organic solvent effects

M. Zama*, D.E. Olins, B. Prescotta and G.J. Thomas, Jra

University of Tennessee - Oak Ridge Graduate School of Biomedical Sciences and the Biology Division, Oak Ridge National Laboratory Oak Ridge, TN 37830, USA aDepartment of Chemistry, Southeastern Massachusetts University North Dartmouth, MA 02747, USA

Received July 20, 1978.

Monomer nucleosomes (v1) from chicken erythrocyte nuclei were examined in aqueous buffers (8>pH>3) and in solvent mixtures (i.e., water and ethanol, ethylene glycol, dioxane, dimethyl sulfoxide, 2-methyl-2,4-pentanediol, polyethylene glycol, sucrose, or urea). Circular dichroism, laser Raman spectroscopy of v1, and the fluoresence of v1 labeled with N-(3-pyrene)maleimide on thiol groups of H3 histone were employed to detect conformational transitions in v1. The results of the pH studies were as follows: 5.5>pH>4.8, suppression of DNA ellipticity and no change of histone {alpha}-helix, 4.6>pH>4.2 an irreversible increase in the B character of DNA, a slight loss of histone {alpha}-helix, and a parallel loss of pyrene excimer fluorescence; 4>pH, aggregation of v1 and protonation of the DNA bases C and A. Results obtained in the studies of v1 in solvent mixtures included the following: sharp conformational transitions that variously involved an increase in the B character of DNA, a slight loss of histone {alpha}-helix, and a loss of pyrene excimer. Different solvents required different concentrations to effect these conformational changes.

*On leave from the National Institute of Radiological Sciences, Chiba-Shi, Japan.


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