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 Qing, K.
Right arrow Articles by Srivastava, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Qing, K.
Right arrow Articles by Srivastava, A.

 Previous Article  |  Next Article 

J Virol, February 1998, p. 1593-1599, Vol. 72, No. 2
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Adeno-Associated Virus Type 2-Mediated Gene Transfer: Correlation of Tyrosine Phosphorylation of the Cellular Single-Stranded D Sequence-Binding Protein with Transgene Expression in Human Cells In Vitro and Murine Tissues In Vivo

Keyun Qing,1,2,3 Benjawan Khuntirat,1,2,3 Cathryn Mah,1,2,3 Dagmar M. Kube,1,2,3 Xu-Shan Wang,1,2,3 Selvarangan Ponnazhagan,1,2,3 Shangzhen Zhou,4 Varavani J. Dwarki,4 Mervin C. Yoder,5 and Arun Srivastava1,2,3,6,*

Department of Microbiology and Immunology,1 Walther Oncology Center,2 Herman B. Wells Center for Pediatric Research and Department of Biochemistry and Molecular Biology,5 and Division of Hematology/Oncology, Department of Medicine,6 Indiana University School of Medicine, and Walther Cancer Institute,3 Indianapolis, Indiana 46202, and Virology Department, Chiron Corporation, Emeryville, California 946084

Received 21 July 1997/Accepted 16 October 1997

Although the adeno-associated virus type 2 (AAV)-based vector system has gained attention as a potentially useful alternative to the more commonly used retroviral and adenoviral vectors for human gene therapy, the single-stranded nature of the viral genome, and consequently the rate-limiting second-strand viral DNA synthesis, significantly affect its transduction efficiency. We have identified a cellular tyrosine phosphoprotein, designated the single-stranded D sequence-binding protein (ssD-BP), which interacts specifically with the D sequence at the 3' end of the AAV genome and may prevent viral second-strand DNA synthesis in HeLa cells (K. Y. Qing et al., Proc. Natl. Acad. Sci. USA 94:10879-10884, 1997). In the present studies, we examined whether the phosphorylation state of the ssD-BP correlates with the ability of AAV to transduce various established and primary cells in vitro and murine tissues in vivo. The efficiencies of transduction of established human cells by a recombinant AAV vector containing the beta -galactosidase reporter gene were 293 > KB > HeLa, which did not correlate with the levels of AAV infectivity. However, the amounts of dephosphorylated ssD-BP which interacted with the minus-strand D probe were also as follows: 293 > KB > HeLa. Predominantly the phosphorylated form of the ssD-BP was detected in cells of the K562 line, a human erythroleukemia cell line, and in CD34+ primary human hematopoietic progenitor cells; consequently, the efficiencies of AAV-mediated transgene expression were significantly lower in these cells. Murine Sca-1+ lin- primary hematopoietic stem/progenitor cells contained predominantly the dephosphorylated form of the ssD-BP, and these cells could be efficiently transduced by AAV vectors. Dephosphorylation of the ssD-BP also correlated with expression of the adenovirus E4orf6 protein, known to induce AAV gene expression. A deletion mutation in the E4orf6 gene resulted in a failure to catalyze dephosphorylation of the ssD-BP. Extracts prepared from mouse brain, heart, liver, lung, and skeletal-muscle tissues, all of which are known to be highly permissive for AAV-mediated transgene expression, contained predominantly the dephosphorylated form of the ssD-BP. Thus, the efficiency of transduction by AAV vectors correlates well with the extent of the dephosphorylation state of the ssD-BP in vitro as well as in vivo. These data suggest that further studies on the cellular gene that encodes the ssD-BP may promote 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: arun_srivastava{at}iucc.iupui.edu.




This article has been cited by other articles:

  • Grimm, D., Pandey, K., Nakai, H., Storm, T. A., Kay, M. A. (2006). Liver Transduction with Recombinant Adeno-Associated Virus Is Primarily Restricted by Capsid Serotype Not Vector Genotype. J. Virol. 80: 426-439 [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]  
  • 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]  
  • Duan, D., Yue, Y., Engelhardt, J. F. (2003). Consequences of DNA-Dependent Protein Kinase Catalytic Subunit Deficiency on Recombinant Adeno-Associated Virus Genome Circularization and Heterodimerization in Muscle Tissue. J. Virol. 77: 4751-4759 [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]  
  • 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]  
  • Zentilin, L., Marcello, A., Giacca, M. (2001). Involvement of Cellular Double-Stranded DNA Break Binding Proteins in Processing of the Recombinant Adeno-Associated Virus Genome. J. Virol. 75: 12279-12287 [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]  
  • Duan, D., Yan, Z., Yue, Y., Ding, W., Engelhardt, J. F. (2001). Enhancement of Muscle Gene Delivery with Pseudotyped Adeno-Associated Virus Type 5 Correlates with Myoblast Differentiation. J. Virol. 75: 7662-7671 [Abstract] [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]  
  • Douar, A.-M., Poulard, K., Stockholm, D., Danos, O. (2001). Intracellular Trafficking of Adeno-Associated Virus Vectors: Routing to the Late Endosomal Compartment and Proteasome Degradation. J. Virol. 75: 1824-1833 [Abstract] [Full Text]  
  • Tattersall, P. (2000). The persistence of alien genomes. Proc. Natl. Acad. Sci. USA 97: 6239-6241 [Full Text]  
  • Bartlett, J. S., Wilcher, R., Samulski, R. J. (2000). Infectious Entry Pathway of Adeno-Associated Virus and Adeno-Associated Virus Vectors. J. Virol. 74: 2777-2785 [Abstract] [Full Text]  
  • Ward, P., Linden, R. M. (2000). A Role for Single-Stranded Templates in Cell-Free Adeno-Associated Virus DNA Replication. J. Virol. 74: 744-754 [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]  
  • Grifman, M., Chen, N. N., Gao, G.-p., Cathomen, T., Wilson, J. M., Weitzman, M. D. (1999). Overexpression of Cyclin A Inhibits Augmentation of Recombinant Adeno-Associated Virus Transduction by the Adenovirus E4orf6 Protein. J. Virol. 73: 10010-10019 [Abstract] [Full Text]  
  • Duan, D., Li, Q., Kao, A. W., Yue, Y., Pessin, J. E., Engelhardt, J. F. (1999). Dynamin Is Required for Recombinant Adeno-Associated Virus Type 2 Infection. J. Virol. 73: 10371-10376 [Abstract] [Full Text]  
  • Yang, J., Zhou, W., Zhang, Y., Zidon, T., Ritchie, T., Engelhardt, J. F. (1999). Concatamerization of Adeno-Associated Virus Circular Genomes Occurs through Intermolecular Recombination. J. Virol. 73: 9468-9477 [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]  
  • Russell, D. W., Kay, M. A. (1999). Adeno-Associated Virus Vectors and Hematology. Blood 94: 864-874 [Full Text]  
  • Smith, D. H., Ward, P., Linden, R. M. (1999). Comparative Characterization of Rep Proteins from the Helper-Dependent Adeno-Associated Virus Type 2 and the Autonomous Goose Parvovirus. J. Virol. 73: 2930-2937 [Abstract] [Full Text]  
  • Daly, T. M., Vogler, C., Levy, B., Haskins, M. E., Sands, M. S. (1999). Neonatal gene transfer leads to widespread correction of pathology in a murine model of lysosomal storage disease. Proc. Natl. Acad. Sci. USA 96: 2296-2300 [Abstract] [Full Text]  
  • Vincent-Lacaze, N., Snyder, R. O., Gluzman, R., Bohl, D., Lagarde, C., Danos, O. (1999). Structure of Adeno-Associated Virus Vector DNA following Transduction of the Skeletal Muscle. J. Virol. 73: 1949-1955 [Abstract] [Full Text]  
  • Mah, C., Qing, K., Khuntirat, B., Ponnazhagan, S., Wang, X.-S., Kube, D. M., Yoder, M. C., Srivastava, A. (1998). Adeno-Associated Virus Type 2-Mediated Gene Transfer: Role of Epidermal Growth Factor Receptor Protein Tyrosine Kinase in Transgene Expression. J. Virol. 72: 9835-9843 [Abstract] [Full Text]  
  • Wang, X.-S., Srivastava, A. (1998). Rescue and Autonomous Replication of Adeno-Associated Virus Type 2 Genomes Containing Rep-Binding Site Mutations in the Viral p5 Promoter. J. Virol. 72: 4811-4818 [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»