Limited proteolysis of rat liver nucleolin by endogenous proteases: effects of polyamines and histones.
Nucleolin is a major nucleolar phosphoprotein and is presumably involved in rDNA transcription and ribosome biosynthesis. This protein is known to be very labile and to be cleaved by endogenous proteases into many small peptides. We found that, when rat liver nucleolar suspension (Nu-1) or nucleolin-rich extract (Nu-2) was incubated under conventional conditions, polyamines and histones interacted with the nucleolin to lead to its preferential degradation to 60 kDa phosphopeptide (p60). The peptide p60 was identified as a peptide containing the N-terminal half of the nucleolin molecule, as judged from peptide-map analysis. Whereas spermine binding to the purified nucleolin was decreased by KCl concentrations above 50 mM, histones (H1, H2B and H3) were able to bind to the nucleolin in the presence of up to 300 mM KCl. A distinct difference between H1 and other histones was found in that H1 could produce p60 from nucleolin in both Nu-1 and Nu-2, whereas H2B and H3 stimulated the degradation of nucleolin to p60 only when Nu-2 was used for the source of nucleolin. A possible relationship between p60 formation and rRNA synthesis is discussed, but its exact role remains to be studied.
Suzuki
T
T
Faculty of Pharmaceutical Sciences, Chiba University, Japan.
Suzuki
N
N
Hosoya
T
T
D000818:Animals; D002466:Cell Nucleolus / Q000378:metabolism; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism*; D006657:Histones / Q000378:metabolism*; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism*; D008297:Male; D009687:Nuclear Proteins / Q000378:metabolism*; D010748:Phosphopeptides / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D016601:RNA-Binding Proteins; D051381:Rats; D017208:Rats, Wistar; D013096:Spermine / Q000378:metabolism*; D000097796:Nucleolin
D010450:Endopeptidases; D006657:Histones; D009687:Nuclear Proteins; D010748:Phosphopeptides; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D013096:Spermine; D000097796:Nucleolin