Preribosomal ribonucleoprotein particles are a major component of a nucleolar matrix fraction.
Biochemical and morphological studies were performed on Novikoff hepatoma ascites cell nucleolar matrix fractions prepared by deoxyribonuclease I digestion and high-molarity salt extractions essentially according to a published method [Berezney, R., & Buchholz, L. A. (1981) Exp. Cell Res. 20, 4995-5002]. The nucleolar matrix fraction was enriched in polypeptides of molecular mass of 28, 37.5, 40, 70, 72, 110 (protein C23), and 160 kDa, compared to the nuclear fraction in which polypeptides of molecular mass of 31, 33.5, 43.5, 46, 50, 56, and 59 kDa were predominant. About one-fourth of the protein, half of the RNA, and less than 4% of the DNA originally present in the nucleoli remained in the matrix fraction. Addition of single agents such as ethylenediaminetetraacetic acid, ribonuclease A, or mercaptoethanol during preparation had no significant effect on the polypeptide composition of the nucleolar matrix fraction. However, the combination of mercaptoethanol and ribonuclease A caused most of the RNA and protein to be removed, including protein C23 and the 160-kDa polypeptide, with polypeptides in the range of Mr 30 000-50 000 remaining. Electron microscopy of nucleolar matrix fractions revealed the presence of particles similar in size to the granular elements of nucleoli. However, when ribonuclease A and mercaptoethanol were included in the procedure, only amorphous material remained. Many proteins of nucleolar preribosomal RNP particles were also associated with the nucleolar matrix fraction. RNA from the nucleolar matrix fraction was enriched in sequences from 18S and 28S ribosomal RNA. These results indicate that preribosomal RNP particles are major constituents of a nucleolar matrix fraction prepared by the deoxyribonuclease I-high-molarity salt method.
D000818:Animals; D002458:Cell Fractionation; D002466:Cell Nucleolus / Q000648:ultrastructure*; D002467:Cell Nucleus / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007700:Kinetics; D008114:Liver Neoplasms, Experimental / Q000032:analysis; D008297:Male; D008854:Microscopy, Electron; D008970:Molecular Weight; D009687:Nuclear Proteins; D000090243:Nucleophosmin; D010455:Peptides / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D012261:Ribonucleoproteins / Q000302:isolation & purification* / Q000378:metabolism
D009687:Nuclear Proteins; D010455:Peptides; D012261:Ribonucleoproteins; D000090243:Nucleophosmin