Summary
Proliferating cell nuclear antigen mRNA levels were determined in human diploid fibroblasts as they progressed through the cell cycle. PCNA message levels were low at G0, gradually increased following entrance into G1, peaked at G1/S, and declined during S phase. PCNA mRNA was determined to have a half life of 12 hours when cells were blocked at the G1/S interface. PCNA protein levels increased two- to three-fold as cells moved from G0 to S phase.
Similar content being viewed by others
References
Almendral, J. M.; Huebsch, D.; Blundell, P. A., et al. Cloning and sequence of the human nuclear protein cyclin: homology with DNA binding proteins. Proc. Natl. Acad. Sci. USA 84:1575–1579; 1987.
Barile, M. Mycoplasma contamination of cell cultures: incidence source, prevention and problems of elimination. In: Kruse, P. F., Jr.; Patterson, M. K., Jr., eds. Tissue culture-methods and applications. New York: Academic Press; 1973:729–735.
Bensadoun, A.; Weinstein, D. Assay of proteins in the presence of interfering materials. Anal. Biochem. 70:241–250; 1976.
Bravo, R.; Celis, J. E. Human proteins sensitive to neoplastic transformation in cultured epithelial and fibroblast cells. Clin. Chem. 28:949–954; 1982.
Bravo, R.; Fey, S. J.; Bellatin, J., et al. Identification of nuclear and of a cytoplasmic polypeptide whose relative proportions are sensitive to changes in the rate of cell proliferation. Exp. Cell Res. 136:311–319; 1981.
Bravo, R.; Frank, R.; Blundell, P. A., et al. Cyclin/PCNA is the auxiliary protein of DNA polymerase-. Nature 326:515–517; 1987.
Bravo, R.; Macdonald-Bravo, H. Changes in the nuclear distribution of cyclin (PCNA) but not its synthesis depend on DNA replication. EMBO J. 4:655–661; 1985.
Bravo, R.; Macdonald-Bravo, H. Existence of two populations of cyclin/proliferating cell nuclear antigen during the cell cycle: association with DNA replication sites. J. Cell Biol. 105:1549–1554; 1987.
Celis, J. E.; Bravo, R. Synthesis of the nuclear protein cyclin in growing, senescent and morphologically transformed human skin fibroblasts. FEBS Lett. 165:21–25; 1984.
Celis, J. E.; Celis, A. Cell cycle-dependent variations in the distribution of the nuclear protein cyclin, proliferating cell nuclear antigen, in cultured cells: subdivision of S-phase. Proc. Natl. Acad. Sci. USA 82:3262–3266; 1985.
Chang, C-D.; Ottavio, L.; Travali, S., et al. Transcriptional and post-transcriptional regulation of proliferating cell nuclear antigen gene. Mol. Cell. Biol. 10:3289–3296; 1990.
Chang, C-D.; Phillips, P.; Lipson, K. E., et al. Senescent human fibroblasts have a post-transcriptional block in the expression of the proliferating cell nuclear antigen gene. J. Biol. Chem. 266:8663–8666; 1991.
Hsu, S-M.;Raine, L.; Fanger, H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. J. Histochem. Cytochem. 29:577–580; 1981.
Jaskulski, D.; DeRiel, J. K.; Mercer, W. E., et al. Inhibition of cellular proliferation by antisense oligodeoxynucleotides to PCNA cyclin. Science 240:1544–1546; 1988.
Kingston, R. E. CsCl purification of RNA from cultured cells. In: Ausubel, F. M.; Brent, R.; Kingston, R. E., et al., eds. Current protocols in molecular biology. New York: Wiley and Sons; 1987:4.2.1–4.2.3.
Laemmli, U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685; 1970.
Liu, Y-C.; Bambara, R. A. Gene expression of PCNA/cyclin in adult tissues and the R3230AC mammary tumor of rat. Biochem. Biophys. Res. Commun. 161:873–882; 1989.
Mathews, M. B. The proliferating cell nuclear antigen, PCNA, a growth regulated DNA replication factor. In: Wang, E.; Warner, H., eds. Growth control during cell aging. Boca Raton: CRC Press; 1989:89–120.
Morris, G. F.; Mathews, M. B. Regulation of proliferating cell nuclear antigen during the cell cycle. J. Biol. Chem. 264:13856–13864; 1989.
Phillips, H. J. Dye exclusion tests for cell viability. In: Kruse, P. F., Jr.; Patterson, M. K., Jr., eds. Tissue culture—methods and applications. New York: Academic Press; 1973:406–408.
Prelich, G.; Stillman, B. Coordinated leading and lagging strand synthesis during SV40 DNA replication in vitro requires PCNA. Cell 53:117–126; 1988.
Sadaie, M. R.; Mathews, M. B. Immunochemical and biochemical analysis of the proliferating cell nuclear antigen (PCNA) in HeLa cells. Exp. Cell Res. 163:423–433; 1986.
Selden, R. F. Analysis of RNA by Northern hybridization. In: Ausubel, F. M., et al., eds. Current protocols in molecular biology. New York: Wiley and Sons; 1987:4.9.1–4.9.4.
Shipman, P. M.; Sabath, D. E.; Fischer, A. H., et al. Cyclin mRNA and protein expression in recombinant interleukin 2 stimulated cloned murine T lymphocytes. J. Cell. Biochem. 38:189–198; 1988.
Tobey, R. A.; Valdez, J. G.; Crissman, H. A. Synchronization of human diploid fibroblasts at multiple stages of the cell cycle. Exp. Cell Res. 179:400–416; 1988.
Towbin, H.; Staehelin, T.; Gordon, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76:4350–4354; 1979.
Zambetti, G.; Dell’Orco, R.; Stein, G., et al. Histone gene expression remains coupled to DNA synthesis during in vitro cellular senescence. Exp. Cell Res. 172:397–403; 1987.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Stewart, C.A., Dell’orco, R.T. Expression of proliferationg cell nuclear antigen during the cell cycle of human diploid fibroblasts. In Vitro Cell Dev Biol - Animal 28, 211–214 (1992). https://doi.org/10.1007/BF02631094
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02631094