Release of in vitro-synthesized poly(A)-containing RNA from isolated rat liver nuclei: characterization of the ribonucleoprotein particles involved.
Nuclei isolated from rat liver were incubated under conditions in which RNA continued to be labeled with precursors for long periods. After 1 hr, during which the rate of RNA synthesis was constant, 25-30% of the newly synthesized RNA was recovered in the postnuclear supernatant. About 3-5% of this fraction was characterized as poly(A)-containing ribonucleoproteins by the following criteria: (i) characteristic elution profile in oligo(dT)-cellulose chromatography; (ii) size distribution of the molecules and their deproteinized RNAs; (iii) buoyant densities in CsCl gradients; (iv) presence of RNaseresistant fragments resembling poly(A)-protein complexes; and (v) identification of the protein components by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The sensitivity of labeling of the RNA synthesized and released from the nuclei to low doses of alpha-amanitin suggests the presence of polymerase II products in the particles. Comparison of the sizes of proteins in these particles with those of free and polysomal messenger ribonucleoproteins as well as with heterogenous nuclear ribonucleoproteins indicates that the released particles contain a protein of 78,000 daltons, which is also present in the other three classes of ribonucleoproteins, presumably at the 3'-poly(A) segments. In addition, a few other proteins, similar in size to those found in the cytoplasmic ribonucleoproteins, were also present in the released particles. It is suggested that proteins associated with heterogenous nuclear RNA are mostly exchanged before or at the time of release of mRNA from the nucleus; the remaining mRNA-associated proteins are added in the cytoplasm, possibly in relation to cytoskeleton attachment, followed by the removal of most of these proteins during polysome formation.
D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D003609:Dactinomycin / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008970:Molecular Weight; D009698:Nucleoproteins / Q000378:metabolism*; D011061:Poly A / Q000096:biosynthesis*; D012313:RNA / Q000096:biosynthesis*; D051381:Rats; D012261:Ribonucleoproteins / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects
D000546:Amanitins; D009698:Nucleoproteins; D012261:Ribonucleoproteins; D003609:Dactinomycin; D011061:Poly A; D012313:RNA