This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Di Pasquale, G.
Right arrow Articles by Stacey, S. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Di Pasquale, G.
Right arrow Articles by Stacey, S. N.

 Previous Article  |  Next Article 

Journal of Virology, October 1998, p. 7916-7925, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Adeno-Associated Virus Rep78 Protein Interacts with Protein Kinase A and Its Homolog PRKX and Inhibits CREB-Dependent Transcriptional Activation

Giovanni Di Pasquale and Simon N. Stacey*

Cancer Research Campaign, Section of Molecular Genetics, Paterson Institute for Cancer Research, Christie Hospital, Manchester M20 4BX, United Kingdom

Received 11 May 1998/Accepted 24 June 1998

Adeno-associated virus (AAV) is a human parvovirus of the genus Dependovirus. AAV replication is largely restricted to cells which are coinfected with a helper virus. In the absence of a helper virus, the AAV genome can integrate into a specific chromosomal site where it remains latent until reactivated by superinfection of the host cell with an appropriate helper virus. Replication functions of AAV have been mapped to the Rep68 and Rep78 gene products. Rep proteins demonstrate DNA binding, endonuclease, and helicase activities and are involved in regulation of transcription from both AAV and heterologous promoters. AAV has been associated with suppression of oncogenicity in a range of viral and nonviral tumors. In this study we sought to identify and study cellular protein targets of AAV Rep, in order to develop a better understanding of the various activities of Rep. We used the yeast two-hybrid system to identify HeLa cell proteins that interact with AAV type 2 Rep78. We isolated several strongly interacting clones which were subsequently identified as PRKX (previously named PKX1), a recently described homolog of the protein kinase A (PKA) catalytic subunit (PKAc). The interaction was confirmed in vitro by using pMal-Rep pull-down assays. The region of Rep78 which interacts was mapped to a C-terminal zinc finger-like domain; Rep68, which lacks this domain, did not interact with PRKX. PRKX demonstrated autophosphorylation and kinase activity towards histone H1 and a PKA oligopeptide target. Autophosphorylation was inhibited by interaction with Rep78. In transfection assays, a PRKX expression vector was shown to be capable of activating CREB-dependent transcription. This activation was suppressed by Rep78 but not by Rep68. Since PRKX is a close homolog of PKAc, we investigated whether Rep78 could interact directly with PKAc. pMal-Rep78 was found to associate with purified PKAc and inhibited its kinase activity. Cotransfection experiments demonstrated that Rep78 could block the activation of CREB by a PKAc expression vector. These experiments suggest that AAV may perturb normal cyclic AMP response pathways in infected cells.

* Corresponding author. Mailing address: Cancer Research Campaign, Section of Molecular Genetics, Paterson Institute for Cancer Research, Christie Hospital, Wilmslow Rd., Manchester M20 4BX, United Kingdom. Phone: 161-446-3186. Fax: 161-446-3109. E-mail: sstacey{at}picr.man.ac.uk.

Journal of Virology, October 1998, p. 7916-7925, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Nash, K., Chen, W., Salganik, M., Muzyczka, N. (2009). Identification of Cellular Proteins That Interact with the Adeno-Associated Virus Rep Protein. J. Virol. 83: 454-469 [Abstract] [Full Text]  
  • Pegoraro, G., Marcello, A., Myers, M. P., Giacca, M. (2006). Regulation of Adeno-Associated Virus DNA Replication by the Cellular TAF-I/Set Complex. J. Virol. 80: 6855-6864 [Abstract] [Full Text]  
  • Needham, P. G., Casper, J. M., Kalman-Maltese, V., Verrill, K., Dignam, J. D., Trempe, J. P. (2006). Adeno-associated virus rep protein-mediated inhibition of transcription of the adenovirus major late promoter in vitro.. J. Virol. 80: 6207-6217 [Abstract] [Full Text]  
  • Berthet, C., Raj, K., Saudan, P., Beard, P. (2005). How adeno-associated virus Rep78 protein arrests cells completely in S phase. Proc. Natl. Acad. Sci. USA 102: 13634-13639 [Abstract] [Full Text]  
  • Li, W., Yu, Z.-X., Kotin, R. M. (2005). Profiles of PrKX Expression in Developmental Mouse Embryo and Human Tissues. J. Histochem. Cytochem. 53: 1003-1009 [Abstract] [Full Text]  
  • James, J. A., Aggarwal, A. K., Linden, R. M., Escalante, C. R. (2004). Structure of adeno-associated virus type 2 Rep40-ADP complex: Insight into nucleotide recognition and catalysis by superfamily 3 helicases. Proc. Natl. Acad. Sci. USA 101: 12455-12460 [Abstract] [Full Text]  
  • Urabe, M., Kogure, K., Kume, A., Sato, Y., Tobita, K., Ozawa, K. (2003). Positive and negative effects of adeno-associated virus Rep on AAVS1-targeted integration. J. Gen. Virol. 84: 2127-2132 [Abstract] [Full Text]  
  • Bossis, I., Chiorini, J. A. (2003). Cloning of an Avian Adeno-Associated Virus (AAAV) and Generation of Recombinant AAAV Particles. J. Virol. 77: 6799-6810 [Abstract] [Full Text]  
  • Junttila, I., Bourette, R. P., Rohrschneider, L. R., Silvennoinen, O. (2003). M-CSF induced differentiation of myeloid precursor cells involves activation of PKC-{delta} and expression of Pkare. J. Leukoc. Biol. 73: 281-288 [Abstract] [Full Text]  
  • Lackner, D. F., Muzyczka, N. (2002). Studies of the Mechanism of Transactivation of the Adeno-Associated Virus p19 Promoter by Rep Protein. J. Virol. 76: 8225-8235 [Abstract] [Full Text]  
  • Li, X., Li, H.-P., Amsler, K., Hyink, D., Wilson, P. D., Burrow, C. R. (2002). PRKX, a phylogenetically and functionally distinct cAMP-dependent protein kinase, activates renal epithelial cell migration and morphogenesis. Proc. Natl. Acad. Sci. USA 99: 9260-9265 [Abstract] [Full Text]  
  • Weger, S., Hammer, E., Heilbronn, R. (2002). Topors, a p53 and topoisomerase I binding protein, interacts with the adeno-associated virus (AAV-2) Rep78/68 proteins and enhances AAV-2 gene expression. J. Gen. Virol. 83: 511-516 [Abstract] [Full Text]  
  • Schmidt, M., Chiorini, J. A., Afione, S., Kotin, R. (2002). Adeno-Associated Virus Type 2 Rep78 Inhibition of PKA and PRKX: Fine Mapping and Analysis of Mechanism. J. Virol. 76: 1033-1042 [Abstract] [Full Text]  
  • Schmidt, M., Afione, S., Kotin, R. M. (2000). Adeno-Associated Virus Type 2 Rep78 Induces Apoptosis through Caspase Activation Independently of p53. J. Virol. 74: 9441-9450 [Abstract] [Full Text]  
  • Mori, Y., Dhepakson, P., Shimamoto, T., Ueda, K., Gomi, Y., Tani, H., Matsuura, Y., Yamanishi, K. (2000). Expression of Human Herpesvirus 6B rep within Infected Cells and Binding of Its Gene Product to the TATA-Binding Protein In Vitro and In Vivo. J. Virol. 74: 6096-6104 [Abstract] [Full Text]  
  • Cathomen, T., Collete, D., Weitzman, M. D. (2000). A Chimeric Protein Containing the N Terminus of the Adeno-Associated Virus Rep Protein Recognizes Its Target Site in an In Vivo Assay. J. Virol. 74: 2372-2382 [Abstract] [Full Text]  
  • Rinaudo, D., Lamartina, S., Roscilli, G., Ciliberto, G., Toniatti, C. (2000). Conditional Site-Specific Integration into Human Chromosome 19 by Using a Ligand-Dependent Chimeric Adeno-Associated Virus/Rep Protein. J. Virol. 74: 281-294 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»