Isolation and characterization of the nuclear matrix in Friend erythroleukemia cells: chromatin and hnRNA interactions with the nuclear matrix.
Nuclear matrices from undifferentiated and differentiated Friend erythroleukemia cells have been obtained by a method which removes DNA in a physiological buffer. These matrices preserved the characteristic topographical distribution of condensed and diffuse "chromatin" regions, as do nuclei in situ or isolated nuclei. Histone H1 was released from the nuclear matrix of undifferentiated cells by 0.3 M KCl; inner core histones were released by 1 M KCl. Nuclear matrix from differentiated cells did not maintain H1, and histone cores were fully released in 0.7 M KCl. KCl removed the core histones as an octameric structure with no evidence of preferential release of any single histone. Electron microscopy of KCl-treated matrix revealed no condensed regions but rather a network of fibrils in the whole DNA-depleted nuclei. When nuclear matrices from both types of cell were exposed to conditions of very low ionic strength, inner core histones and condensed regions remained. These observations support the contention that inner core histones are bound to matrix through natural ionic bonds or saline-labile elements, and that these interactions are implicated in chromatin condensation. hnRNA remained undegraded and tenaciously associated to the matrix fibrils, and was released only by chemical means which, by breaking hydrophobic and hydrogen bonds, produced matrix lysis. Very few nonhistone proteins were released upon complete digestion of DNA from either type of nuclei. The remaining nonhistone proteins represent a large number of species of which the majority may be matrix components. The molecular architecture in both condensed and diffuse regions of interphase nuclei appears to be constructed of two distinct kinds of fibers; the thicker chromatin fibers are interwoven with the thinner matrix fibers. The latter are formed by a heteropolymer of many different proteins.
D000818:Animals; D002454:Cell Differentiation; D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus / Q000032:analysis / Q000648:ultrastructure*; D055598:Chemical Phenomena; D002621:Chemistry; D002843:Chromatin / Q000032:analysis*; D003851:Deoxyribonucleases / Q000494:pharmacology; D006657:Histones / Q000032:analysis*; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D009994:Osmolar Concentration; D011506:Proteins / Q000032:analysis; D012332:RNA, Heterogeneous Nuclear / Q000032:analysis*
D002843:Chromatin; D006657:Histones; D011506:Proteins; D012332:RNA, Heterogeneous Nuclear; D003851:Deoxyribonucleases