Immunochemical and biochemical analysis of the proliferating cell nuclear antigen (PCNA) in HeLa cells.
PCNA, also known a cyclin, a protein of molecular weight (MW) 35 000, accumulates in the nuclei of dividing and transformed cells and reacts with autoantibodies from certain lupus patients. Using an indirect immunofluorescence technique, we show that lupus sera containing anti-PCNA antibodies reveal a heterogeneous nuclear fluorescence pattern upon reaction with asynchronous HeLa cells, whereas with synchronized cells a sequence of distinct patterns is disclosed on progression through the cell cycle. Cell-free translation of HeLa cell mRNA followed by immunoprecipitation with anti-PCNA sera shows a single protein with the same apparent MW as PCNA labelled in vivo, suggesting that PCNA is not derived from a larger precursor protein and not grossly modified by post-translational events. However, a group of at least nine nuclear polypeptides ranging in MW from about 12 000 to 110 000 are recognized by immunoblotting with anti-PCNA sera, indicating either that additional antigenic sites are produced on denaturation of native proteins or that additional autoantibodies are present in these sera. We also show that PCNA and several of these polypeptides are associated with nuclear structures containing chromatin.
Sadaie
M R
MR
Mathews
M B
MB
D000941:Antigens / Q000032:analysis*; D001324:Autoantigens / Q000032:analysis*; D002453:Cell Cycle; D006367:HeLa Cells / Q000166:cytology; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D009698:Nucleoproteins / Q000032:analysis* / Q000096:biosynthesis; D018809:Proliferating Cell Nuclear Antigen; D014176:Protein Biosynthesis
D000941:Antigens; D001324:Autoantigens; D009698:Nucleoproteins; D018809:Proliferating Cell Nuclear Antigen