Isolation from Friend erythroleukemia cells of an RNase-sensitive nuclear matrix fibril fraction containing hnRNA and snRNA.
Chromatin-depleted nuclei (CDN) were prepared from Friend erythroleukemia cell nuclei by partial digestion with DNase I and extraction of the chromatin by 2 mM EDTA as described in the preceding paper (Long and Ochs, 1983. Biol. Cell 48, 99-108). These structures contained dense networks of matrix fibrils surrounded by distinct laminae but no morphologically distinct residual nucleoli. CDN disrupted by gentle shearing or 1 microgram/ml RNase were fractionated into laminae and matrix fibrils by differential centrifugation. Protein composition of the lamina fraction was dominated by two prominent lamina proteins that were not detectable in the matrix fraction. Mild RNase treatment led to a conversion of the fibrous network to a particulate morphology while mild shearing resulted in an apparently unaltered fibril fraction. The matrix fibril fractions contained hnRNP proteins and the snRNAs. These results suggest that EDTA-prepared CDN may provide a system for studying snRNP-hnRNP interactions and hnRNP processing that is less complex than intact nuclei.
D000818:Animals; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000648:ultrastructure*; D002843:Chromatin / Q000648:ultrastructure; D007700:Kinetics; D007942:Leukemia, Experimental / Q000648:ultrastructure*; D051379:Mice; D008854:Microscopy, Electron; D012313:RNA / Q000302:isolation & purification*; D012332:RNA, Heterogeneous Nuclear / Q000302:isolation & purification*; D012342:RNA, Small Nuclear; D012260:Ribonucleases / Q000378:metabolism
D002843:Chromatin; D012332:RNA, Heterogeneous Nuclear; D012342:RNA, Small Nuclear; D012313:RNA; D012260:Ribonucleases