Circular dichroic studies of the DNA and RNA of nucleoli.
Circular dichroism (CD) in the 240-300-nm region was used to study the conformation of DNA and RNA complexed with proteins in isolated nucleoli form HeLa cells. Deoxyribonuclease or ribonuclease digestion was employed to obtain (1) the individual CD spectra of nucleolar DNA or RNA in complex form with proteins, or in free form; and (2) the experimental CD baseline correction to exclude contributions from nonnucleic acid sources such as light scattering  artifacts and proteins. The CD spectrum of nucleolar DNA in DNA-protein complexes was highly reduced in ellipticity in comparison with protein-free DNA. It showed a positive peak at 283 nm with a molar ellipticity [theta]283 = 1200 deg cm2 dmol-1 and a crossover at 262 nm. Addition of sodium dodecylsulfate shifted the peak to 276 nm with [theta]276 8000 deg cm2 dmol-1 and a crossover at 254 nm. The CD spectrum of nucleolar RNA in RNA-protein complexes was also reduced in comparison with protein-free RNA, showing a peak at 269 nm ([theta]269 = 6900 deg cm2 dmol-1), and a crossover at 250 nm. Addition of sodium dodecyl sulfate shifted the peak to 265 nm with [theta]265 = 18 000 deg cm2 dmol-1 and a crossover at 246 nm. The low ellipticity of both nucleolar DNA and RNA when complexed with proteins was increased by treatment with sodium chloride, urea, or heparin. This suggests that some ionic, hydrophobic, and hydrogen bondings are involved in the nucleic acid-protein interaction in nucleolar chromatin similar to that observed in nuclear chromatin.
D000546:Amanitins; D001665:Binding Sites; D002458:Cell Fractionation; D004273:DNA, Neoplasm / Q000096:biosynthesis; D003851:Deoxyribonucleases; D003852:Deoxyribonucleoproteins; D006367:HeLa Cells / Q000378:metabolism; D006493:Heparin; D009690:Nucleic Acid Conformation; D009994:Osmolar Concentration; D011189:Potassium Chloride; D012334:RNA, Neoplasm / Q000096:biosynthesis; D012260:Ribonucleases; D012261:Ribonucleoproteins
D000546:Amanitins; D004273:DNA, Neoplasm; D003852:Deoxyribonucleoproteins; D012334:RNA, Neoplasm; D012261:Ribonucleoproteins; D011189:Potassium Chloride; D006493:Heparin; D003851:Deoxyribonucleases; D012260:Ribonucleases