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 Mah, C.
Right arrow Articles by Srivastava, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Mah, C.
Right arrow Articles by Srivastava, A.

 Previous Article  |  Next Article 

Journal of Virology, December 1998, p. 9835-9843, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Adeno-Associated Virus Type 2-Mediated Gene Transfer: Role of Epidermal Growth Factor Receptor Protein Tyrosine Kinase in Transgene Expression

Cathryn Mah,1,2,3 Keyun Qing,1,2,3 Benjawan Khuntirat,1,2,3 Selvarangan Ponnazhagan,1,2,3 Xu-Shan Wang,1,2,3 Dagmar M. Kube,1,2,3 Mervin C. Yoder,4 and Arun Srivastava1,2,3,5,*

Department of Microbiology and Immunology,1 Walther Oncology Center,2 Herman B. Wells Center for Pediatric Research and Department of Biochemistry and Molecular Biology,4 Division of Hematology/Oncology, Department of Medicine, Indiana University School of Medicine,5 and Walther Cancer Institute,3 Indianapolis, Indiana 46202

Received 20 March 1998/Accepted 14 September 1998

Adeno-associated virus type 2 (AAV), a single-stranded, DNA-containing, nonpathogenic human parvovirus, has gained attention as a potentially useful vector for human gene therapy. However, the transduction efficiency of AAV vectors varies greatly in different cells and tissues in vitro and in vivo. We have recently documented that a cellular tyrosine phosphoprotein, designated the single-stranded D-sequence-binding protein (ssD-BP), plays an important role in AAV-mediated transgene expression (K. Y. Qing et al., Proc. Natl. Acad. Sci. USA 94:10879-10884, 1997) and that a strong correlation exists between the phosphorylation state of the ssD-BP and AAV transduction efficiency in vitro as well as in vivo (K. Y. Qing et al., J. Virol. 72:1593-1599, 1998). In this report, we document that treatment of cells with specific inhibitors of the epidermal growth factor receptor protein tyrosine kinase (EGF-R PTK) activity, such as tyrphostin, leads to significant augmentation of AAV transduction efficiency, and phosphorylation of the ssD-BP is mediated by the EGF-R PTK. Treatment of cells with EGF results in phosphorylation of the ssD-BP, whereas treatment with tyrphostin causes dephosphorylation of the ssD-BP and consequently leads to increased expression of the transgene. Furthermore, AAV transduction efficiency inversely correlates with expression of the EGF-R in different cell types, and stable transfection of the EGF-R cDNA causes phosphorylation of the ssD-BP, leading to significant inhibition in AAV-mediated transgene expression which can be overcome by the tyrphostin treatment. These data suggest that the PTK activity of the EGF-R is a crucial determinant in the life cycle of AAV and that further studies on the interaction between the EGF-R and the ssD-BP may yield new insights not only into its role in the host cell but also in the successful use of AAV vectors in human gene therapy.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Indiana University School of Medicine, 635 Barnhill Dr., Medical Science Building, Room 257, Indianapolis, IN 46202-5120. Phone: (317) 274-2194. Fax: (317) 274-4090. E-mail: asrivast{at}iupui.edu.

Journal of Virology, December 1998, p. 9835-9843, Vol. 72, No. 12
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Zhong, L., Li, B., Mah, C. S., Govindasamy, L., Agbandje-McKenna, M., Cooper, M., Herzog, R. W., Zolotukhin, I., Warrington, K. H. Jr., Weigel-Van Aken, K. A., Hobbs, J. A., Zolotukhin, S., Muzyczka, N., Srivastava, A. (2008). Next generation of adeno-associated virus 2 vectors: Point mutations in tyrosines lead to high-efficiency transduction at lower doses. Proc. Natl. Acad. Sci. USA 105: 7827-7832 [Abstract] [Full Text]  
  • Han, Z., Zhong, L., Srivastava, A., Stacpoole, P. W. (2008). Pyruvate Dehydrogenase Complex Deficiency Caused by Ubiquitination and Proteasome-mediated Degradation of the E1 Subunit. J. Biol. Chem. 283: 237-243 [Abstract] [Full Text]  
  • Nathwani, A. C., Gray, J. T., Ng, C. Y. C., Zhou, J., Spence, Y., Waddington, S. N., Tuddenham, E. G. D., Kemball-Cook, G., McIntosh, J., Boon-Spijker, M., Mertens, K., Davidoff, A. M. (2006). Self-complementary adeno-associated virus vectors containing a novel liver-specific human factor IX expression cassette enable highly efficient transduction of murine and nonhuman primate liver. Blood 107: 2653-2661 [Abstract] [Full Text]  
  • Sugano, E., Tomita, H., Ishiguro, S.-i., Abe, T., Tamai, M. (2005). Establishment of Effective Methods for Transducing Genes into Iris Pigment Epithelial Cells by Using Adeno-associated Virus Type 2. IOVS 46: 3341-3348 [Abstract] [Full Text]  
  • Zhong, L., Qing, K., Si, Y., Chen, L., Tan, M., Srivastava, A. (2004). Heat-shock Treatment-mediated Increase in Transduction by Recombinant Adeno-associated Virus 2 Vectors Is Independent of the Cellular Heat-shock Protein 90. J. Biol. Chem. 279: 12714-12723 [Abstract] [Full Text]  
  • Davidoff, A. M., Ng, C. Y. C., Zhou, J., Spence, Y., Nathwani, A. C. (2003). Sex significantly influences transduction of murine liver by recombinant adeno-associated viral vectors through an androgen-dependent pathway. Blood 102: 480-488 [Abstract] [Full Text]  
  • Smith, A. D., Collaco, R. F., Trempe, J. P. (2003). Enhancement of Recombinant Adeno-Associated Virus Type 2-Mediated Transgene Expression in a Lung Epithelial Cell Line by Inhibition of the Epidermal Growth Factor Receptor. J. Virol. 77: 6394-6404 [Abstract] [Full Text]  
  • Qing, K., Li, W., Zhong, L., Tan, M., Hansen, J., Weigel-Kelley, K. A., Chen, L., Yoder, M. C., Srivastava, A. (2003). Adeno-Associated Virus Type 2-Mediated Gene Transfer: Role of Cellular T-Cell Protein Tyrosine Phosphatase in Transgene Expression in Established Cell Lines In Vitro and Transgenic Mice In Vivo. J. Virol. 77: 2741-2746 [Abstract] [Full Text]  
  • Ponnazhagan, S., Mahendra, G., Kumar, S., Thompson, J. A., Castillas, M. Jr. (2002). Conjugate-Based Targeting of Recombinant Adeno-Associated Virus Type 2 Vectors by Using Avidin-Linked Ligands. J. Virol. 76: 12900-12907 [Abstract] [Full Text]  
  • Wendtner, C.-M., Kofler, D. M., Theiss, H. D., Kurzeder, C., Buhmann, R., Schweighofer, C., Perabo, L., Danhauser-Riedl, S., Baumert, J., Hiddemann, W., Hallek, M., Buning, H. (2002). Efficient gene transfer of CD40 ligand into primary B-CLL cells using recombinant adeno-associated virus (rAAV) vectors. Blood 100: 1655-1661 [Abstract] [Full Text]  
  • Qing, K., Hansen, J., Weigel-Kelley, K. A., Tan, M., Zhou, S., Srivastava, A. (2001). Adeno-Associated Virus Type 2-Mediated Gene Transfer: Role of Cellular FKBP52 Protein in Transgene Expression. J. Virol. 75: 8968-8976 [Abstract] [Full Text]  
  • Ponnazhagan, S., Mahendra, G., Curiel, D. T., Shaw, D. R. (2001). Adeno-Associated Virus Type 2-Mediated Transduction of Human Monocyte-Derived Dendritic Cells: Implications for Ex Vivo Immunotherapy. J. Virol. 75: 9493-9501 [Abstract] [Full Text]  
  • Ponnazhagan, S., Curiel, D. T., Shaw, D. R., Alvarez, R. D., Siegal, G. P. (2001). Adeno-associated Virus for Cancer Gene Therapy. Cancer Res. 61: 6313-6321 [Full Text]  
  • Hansen, J., Qing, K., Srivastava, A. (2001). Adeno-Associated Virus Type 2-Mediated Gene Transfer: Altered Endocytic Processing Enhances Transduction Efficiency in Murine Fibroblasts. J. Virol. 75: 4080-4090 [Abstract] [Full Text]  
  • Hansen, J., Qing, K., Kwon, H.-J., Mah, C., Srivastava, A. (2000). Impaired Intracellular Trafficking of Adeno-Associated Virus Type 2 Vectors Limits Efficient Transduction of Murine Fibroblasts. J. Virol. 74: 992-996 [Abstract] [Full Text]  
  • Wang, D., Parrish, C. R. (1999). A Heterogeneous Nuclear Ribonucleoprotein A/B-Related Protein Binds to Single-Stranded DNA near the 5' End or within the Genome of Feline Parvovirus and Can Modify Virus Replication. J. Virol. 73: 7761-7768 [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»