Non-histone proteins in fractionated chromatin before and after DNAse II treatment.
Chromatin from spleen cells of normal, non-immunized mice and from mice 3 days after immunization with human immunoglobulin G was fractionated at increasing salt concentrations into three fractions: 0.35 M NaCl-soluble, 2 M NaCl-soluble and a residual fraction, dissociated in 2 M NaCl/5 M urea. The residual fraction of chromatin, homogeneous by ultracentrifugation and containing only 25% of the total chromatin DNA, was associated with proteins strongly labeled with [3H]tryptophan, [3H]methionine and [3H]leucine. This fraction was more sensitive to DNAase II treatment than was native, non-fractionated chromatin and it contained approx. 40% Mg2+-soluble DNA sequences. The template activity of the residual fraction was 6--7-times higher than that of non-fractionated chromatin. Fraction A, characteristic for non-immunized spleen cells, was present in three chromatin fractions and after DNAase II treatment it remained only in the residual fraction, which suggests that this fraction is associated with genes non-transcribed in non-immunized mice. Fractions I and B1 were found mainly in the residual fraction, and only in smaller amounts in the 0.35 M NaCl-soluble fraction. After DNAase II treatment, fractions I and B1 in chromatin from immunized mice disappeared, which suggests that these fractions may be associated with active transcribed sequences during the immune reaction.
D000818:Animals; D000917:Antibody Formation; D002843:Chromatin / Q000302:isolation & purification / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000096:biosynthesis*; D004247:DNA / Q000378:metabolism; D003851:Deoxyribonucleases; D004706:Endodeoxyribonucleases; D004720:Endonucleases; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D012319:RNA Polymerase II / Q000494:pharmacology; D013154:Spleen / Q000378:metabolism*; D013698:Templates, Genetic; D014158:Transcription, Genetic
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D007074:Immunoglobulin G; D004247:DNA; D012319:RNA Polymerase II; D003851:Deoxyribonucleases; D004706:Endodeoxyribonucleases; D004720:Endonucleases; C019329:deoxyribonuclease II