Effects of various salts and pH on the stability of the nucleosome in chromatin fragments.
The stability of nucleosomes in long chromatin fragments was observed by differential scanning calorimetry over a wide range of solution conditions. The thermal denaturation of chromatin was characterized in general as three major transitions, although the process clearly is more complex. The three major transitions were (1) denaturation of the nucleosome, (2) base unstacking of DNA in the resulting denatured nucleoprotein, and (3) base unstacking of naked DNA. In very low salt concentrations (e.g., 2 mM sodium cacodylate), these three processes were essentially coincident (near 76 degrees C), but in medium salt concentrations (e.g., 100 mM NaCl) the nucleosome denaturation occurred first at about 69 degrees C and then base unstacking occurred at 85 degrees C. As [NaCl] was increased, all three processes were resolved with the observation of increasing amounts of naked DNA being melted, until at 2000 mM NaCl the calorimetric profile showed mainly the melting of DNA. The transition temperature for nucleosome denaturation decreased from 76 to 63 degrees C as the salt concentration increased from 1 to 600 mM. Destabilization of the nucleosome by increasing [NaCl] was also evident above 100 mM as a decrease in enthalpic change attributable to nucleosome denaturation. Similarly, as [NaCl] was increased above 100 mM, less and less denatured nucleoprotein was evident as more and more of the DNA melted as naked DNA. The fatty acid salts, sodium valerate and sodium caproate, destabilized the nucleosome but not the denatured nucleoprotein that resulted from the collapse of the nucleosome. In the series acetate, butyrate, valerate, caproate, it was clear that destabilization of the nucleosome increased as hydrophobicity (chain length) increased.(ABSTRACT TRUNCATED AT 250 WORDS)
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Department of Molecular and Cell Biology, University of California, Berkeley 94720.
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D002152:Calorimetry, Differential Scanning; D002843:Chromatin / Q000737:chemistry* / Q000187:drug effects; D004273:DNA, Neoplasm / Q000737:chemistry* / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions; D015636:Magnesium Chloride / Q000494:pharmacology; D009994:Osmolar Concentration; D010421:Pentanoic Acids / Q000494:pharmacology; D012492:Salts / Q000494:pharmacology*; D012965:Sodium Chloride / Q000494:pharmacology*; D013095:Spermidine / Q000494:pharmacology
D002843:Chromatin; D004273:DNA, Neoplasm; D007477:Ions; D010421:Pentanoic Acids; D012492:Salts; D015636:Magnesium Chloride; D012965:Sodium Chloride; C038780:n-pentanoic acid; D013095:Spermidine