Original Article

Formation of a Complex between Nucleolin and Replication Protein A after Cell Stress Prevents Initiation of DNA Replication

Correspondence to: James A. Borowiec, Department of Biochemistry, New York University School of Medicine, 550 First Avenue, New York, NY 10016. Tel:(212) 263-8453 Fax:(212) 263-8166 E-mail:james.borowiec{at}med.nyu.edu.

We used a biochemical screen to identify nucleolin, a key factor in ribosome biogenesis, as a high-affinity binding partner for the heterotrimeric human replication protein A (hRPA). Binding studies in vitro demonstrated that the two proteins physically interact, with nucleolin using an unusual contact with the small hRPA subunit. Nucleolin significantly inhibited both simian virus 40 (SV-40) origin unwinding and SV-40 DNA replication in vitro, likely by nucleolin preventing hRPA from productive interaction with the SV-40 initiation complex. In vivo, use of epifluorescence and confocal microscopy showed that heat shock caused a dramatic redistribution of nucleolin from the nucleolus to the nucleoplasm. Nucleolin relocalization was concomitant with a tenfold increase in nucleolin–hRPA complex formation. The relocalized nucleolin significantly overlapped with the position of hRPA, but only poorly with sites of ongoing DNA synthesis. We suggest that the induced nucleolin–hRPA interaction signifies a novel mechanism that represses chromosomal replication after cell stress.

Key Words: replication protein A, nucleolin, nucleolus, SV-40, heat shock

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Calle, A., Ugrinova, I., Epstein, A. L., Bouvet, P., Diaz, J.-J., Greco, A. (2008). Nucleolin Is Required for an Efficient Herpes Simplex Virus Type 1 Infection. J. Virol. 82: 4762-4773 [Abstract] [Full Text]  
• Yang, L., Reece, J. M., Cho, J., Bortner, C. D., Shears, S. B. (2008). The Nucleolus Exhibits an Osmotically Regulated Gatekeeping Activity That Controls the Spatial Dynamics and Functions of Nucleolin. J. Biol. Chem. 283: 11823-11831 [Abstract] [Full Text]  
• Fanning, E., Klimovich, V., Nager, A. R. (2006). A dynamic model for replication protein A (RPA) function in DNA processing pathways. Nucleic Acids Res 34: 4126-4137 [Abstract] [Full Text]  
• Christov, C. P., Gardiner, T. J., Szuts, D., Krude, T. (2006). Functional Requirement of Noncoding Y RNAs for Human Chromosomal DNA Replication.. Mol. Cell. Biol. 26: 6993-7004 [Abstract] [Full Text]  
• Girvan, A. C., Teng, Y., Casson, L. K., Thomas, S. D., Juliger, S., Ball, M. W., Klein, J. B., Pierce, W. M. Jr., Barve, S. S., Bates, P. J. (2006). AGRO100 inhibits activation of nuclear factor-{kappa}B (NF-{kappa}B) by forming a complex with NF-{kappa}B essential modulator (NEMO) and nucleolin.. Molecular Cancer Therapeutics 5: 1790-1799 [Abstract] [Full Text]  
• Wu, Y.-L., Dudognon, C., Nguyen, E., Hillion, J., Pendino, F., Tarkanyi, I., Aradi, J., Lanotte, M., Tong, J.-H., Chen, G.-Q., Segal-Bendirdjian, E. (2006). Immunodetection of human telomerase reverse-transcriptase (hTERT) re-appraised: nucleolin and telomerase cross paths. J. Cell Sci. 119: 2797-2806 [Abstract] [Full Text]  
• Kim, K., Dimitrova, D. D., Carta, K. M., Saxena, A., Daras, M., Borowiec, J. A. (2005). Novel Checkpoint Response to Genotoxic Stress Mediated by Nucleolin-Replication Protein A Complex Formation. Mol. Cell. Biol. 25: 2463-2474 [Abstract] [Full Text]  
• Weisshart, K., Pestryakov, P., Smith, R. W. P., Hartmann, H., Kremmer, E., Lavrik, O., Nasheuer, H.-P. (2004). Coordinated Regulation of Replication Protein A Activities by Its Subunits p14 and p32. J. Biol. Chem. 279: 35368-35376 [Abstract] [Full Text]  
• Eilbracht, J., Reichenzeller, M., Hergt, M., Schnolzer, M., Heid, H., Stohr, M., Franke, W. W., Schmidt-Zachmann, M. S. (2004). NO66, a Highly Conserved Dual Location Protein in the Nucleolus and in a Special Type of Synchronously Replicating Chromatin. Mol. Biol. Cell 15: 1816-1832 [Abstract] [Full Text]  
• Vassin, V. M., Wold, M. S., Borowiec, J. A. (2004). Replication Protein A (RPA) Phosphorylation Prevents RPA Association with Replication Centers. Mol. Cell. Biol. 24: 1930-1943 [Abstract] [Full Text]  
• Mi, Y., Thomas, S. D., Xu, X., Casson, L. K., Miller, D. M., Bates, P. J. (2003). Apoptosis in Leukemia Cells Is Accompanied by Alterations in the Levels and Localization of Nucleolin. J. Biol. Chem. 278: 8572-8579 [Abstract] [Full Text]  
• Daniely, Y., Dimitrova, D. D., Borowiec, J. A. (2002). Stress-Dependent Nucleolin Mobilization Mediated by p53-Nucleolin Complex Formation. Mol. Cell. Biol. 22: 6014-6022 [Abstract] [Full Text]  
• Denegri, M., Moralli, D., Rocchi, M., Biggiogera, M., Raimondi, E., Cobianchi, F., De Carli, L., Riva, S., Biamonti, G. (2002). Human Chromosomes 9, 12, and 15 Contain the Nucleation Sites of Stress-Induced Nuclear Bodies. Mol. Biol. Cell 13: 2069-2079 [Abstract] [Full Text]  
• Xu, X., Hamhouyia, F., Thomas, S. D., Burke, T. J., Girvan, A. C., McGregor, W. G., Trent, J. O., Miller, D. M., Bates, P. J. (2001). Inhibition of DNA Replication and Induction of S Phase Cell Cycle Arrest by G-rich Oligonucleotides. J. Biol. Chem. 276: 43221-43230 [Abstract] [Full Text]  
• Edwards, T. K., Saleem, A., Shaman, J. A., Dennis, T., Gerigk, C., Oliveros, E., Gartenberg, M. R., Rubin, E. H. (2000). Role for Nucleolin/Nsr1 in the Cellular Localization of Topoisomerase I. J. Biol. Chem. 275: 36181-36188 [Abstract] [Full Text]  
• Maga, G., Frouin, I., Spadari, S., Hubscher, U. (2001). Replication Protein A as a "Fidelity Clamp" for DNA Polymerase alpha. J. Biol. Chem. 276: 18235-18242 [Abstract] [Full Text]  
• Wang, Y., Guan, J., Wang, H., Wang, Y., Leeper, D., Iliakis, G. (2001). Regulation of DNA Replication after Heat Shock by Replication Protein A-Nucleolin Interactions. J. Biol. Chem. 276: 20579-20588 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents