Isolation and characterization of rat liver free and membrane-bound polysomal messenger ribonucleoprotein particles.
Rat liver mRNA-labeled free and membrane-bound polysomes uncontaminated with nuclear or cytoplasmic ribonucleoprotein particles were dissociated with EDTA and the released messenger ribonucleoprotein particles isolated using an oligo(dT)-cellulose column. Seventy percent of the labeled mRNA applied bound to the column. Binding of mRNP was dependent on the presence of a poly(A) segment in the RNA. The nonbound material contained most of the ribosomal subunits and also messenger-sized poly(A-)RNA molecules associated with protein. The free mRNP fraction bound to the column, washed with 250 mM NaCl, and subsequently eluted contained seven proteins ranging in molecular weight from 52 000 to 138 000 only one of which was found in the fraction not bound to the column. Furthermore, these proteins were shown to have a higher affinity for poly(A+)RNA as compared with rRNA. The membrane-bound mRNP contained five proteins, four of which were identical with those associated with free mRNP. Membrane-bound mRNP were disrupted at a lower salt concentration than the free. The proteins found associated with free and membrane-bound polysomal mRNP appeared to be clustered in the poly(A) region of the molecule. The implications of these findings are discussed.
D000818:Animals; D002462:Cell Membrane; D008099:Liver / Q000032:analysis*; D008297:Male; D008970:Molecular Weight; D009698:Nucleoproteins / Q000302:isolation & purification; D011061:Poly A / Q000032:analysis; D011132:Polyribosomes / Q000032:analysis*; D012333:RNA, Messenger / Q000302:isolation & purification; D051381:Rats; D012261:Ribonucleoproteins / Q000302:isolation & purification
D009698:Nucleoproteins; D012333:RNA, Messenger; D012261:Ribonucleoproteins; D011061:Poly A