Biochemical characterization of chromatin fractions isolated from induced and uninduced Friend erythroleukemia cells.
Chromatin fractions from Friend erythroleukemia cells after induction of differentiation by dimethylsulfoxide (DMSO) were compared in their biochemical characteristics to fractions from uninduced cells. Fractions were prepared by extracting chromatin from nuclei after mild micrococcal nuclease treatment with increasing concentrations of NaCl according to Sanders. This procedure has been found to release chromatin containing hyperacetylated histones preferentially. The fractions obtained by this procedure were analysed in respect to the amount of chromatin released, the amount of histone H1, the degree of acetylation of histone H4, the presence of non-histone proteins and the concentration of transcribed and non-transcribed sequences. It was found that the fractions differ in the amount of histone H1 present, in several non-histone proteins and in the acetylation of histone H4, regardless whether induced or uninduced cells were analysed. The distribution of transcribed sequences versus non-transcribed sequences among the fractions was the same, demonstrating that this fractionation procedure, although leading to fractions with biochemical differences, is not able to discriminate functional states of chromatin and that the biochemical characteristics of the fractions may be common to both, active as well as inactive states of chromatin.
Knosp
O
O
Institut für Medizinische Chemie und Biochemie der Universität, Innsbruck, Austria.
Redl
B
B
Puschendorf
B
B
D000107:Acetylation; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D002843:Chromatin / Q000032:analysis*; D004247:DNA / Q000032:analysis; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005622:Friend murine leukemia virus; D005914:Globins / Q000235:genetics; D006657:Histones / Q000302:isolation & purification / Q000378:metabolism*; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism*; D051379:Mice; D008836:Micrococcal Nuclease; D011499:Protein Processing, Post-Translational; D014444:Tyrosine Transaminase / Q000235:genetics
D002843:Chromatin; D006657:Histones; D005914:Globins; D004247:DNA; D014444:Tyrosine Transaminase; D008836:Micrococcal Nuclease; D004121:Dimethyl Sulfoxide