title: Nuclear matrix-like filaments and fibrogranular complexes form through the rearrangement of specific nuclear ribonucleoproteins.
abstract: The behavior of nuclear pre-mRNA-binding proteins after their nuclease and/or salt-induced release from RNA was investigated. After RNase digestion or salt extraction, two proteins that initially exist as tetramers (A2)(3)B1 in isolated heterogeneous nuclear ribonucleoprotein (hnRNP) complexes quantitatively reassociated to form regular helical filaments ranging in length from 100 nm to >10 microm. In highly magnified preparations prepared for scanning transmission electron microscopy, single filaments have diameters near 18 nm. In conventional negatively stained preparations viewed at low magnification, the diameters of the thinnest filaments range from 7 to 10 nm. At protein concentrations of >0.1 mg/ml, the filaments rapidly aggregated to form thicker filamentous networks that look like the fibrogranular structures termed the "nuclear matrix." Like the residual material seen in nuclear matrix preparations, the hnRNP filaments were insoluble in 2 M NaCl. Filament formation is associated with, and may be dependent on, disulfide bridge formation between the hnRNP proteins. The reducing agent 2-mercaptoethanol significantly attenuates filament assembly, and the residual material that forms is ultrastructurally distinct from the 7- to 10-nm fibers. In addition to the protein rearrangement leading to filament formation, nearly one-third of the protein present in chromatin-clarified nuclear extracts was converted to salt-insoluble material within 1 min of digestion with RNase. These observations are consistent with the possibility that the residual material termed the nuclear matrix may be enriched in, if not formed by, denatured proteins that function in pre-mRNA packaging, processing, and transport.
authors.affiliation: Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235, USA.
authors.forename: J H
authors.initials: JH
authors.lastname: Tan
authors.forename: J C
authors.initials: JC
authors.lastname: Wooley
authors.forename: W M
authors.initials: WM
authors.lastname: LeStourgeon
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure*; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000648:ultrastructure*; D004220:Disulfides; D006801:Humans; D051379:Mice; D051381:Rats; D012260:Ribonucleases / Q000737:chemistry / Q000378:metabolism; D012261:Ribonucleoproteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D012965:Sodium Chloride / Q000737:chemistry; D012995:Solubility
chemical_list: D004220:Disulfides; D012261:Ribonucleoproteins; D012965:Sodium Chloride; D012260:Ribonucleases