Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

The HIV Tat protein transduction domain improves the biodistribution of β-glucuronidase expressed from recombinant viral vectors

Nature Biotechnology volume 19, pages 640–644 (2001)Cite this article

Abstract

Treatment of inherited genetic diseases of the brain remains an intractable problem. Methods to improve the distribution of enzymes that are injected or expressed from transduced cells will be required for many human brain therapies. Recent studies showed that a peptide, the protein transduction domain (PTD) from HIV Tat, could improve the distribution of cytoplasmic reporter proteins when administered systemically as fusion proteins or cross-linked chimeras. The utility of this motif for noncytoplasmic proteins has not been determined. Here, we tested how the Tat motif affected uptake and biodistribution of the lysosomal enzyme β-glucuronidase, the protein deficient in the disease mucopolysaccharidosis VII, when expressed from viral vectors. The Tat motif allowed for mannose-6-phosphate (M6P) independent uptake in vitro and significantly increased the distribution of β-glucuronidase secreted from transduced cells after intravenous or direct brain injection in mice of recombinant vectors. Thus, enzymes modified to contain protein transduction motifs may represent a general strategy for improving the distribution of secreted proteins following in vivo gene transfer.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: β-Glucuronidase–Tat expression vectors and recombinant protein uptake.
Figure 2: Activity of eGFP and β-glucuronidase in sections of murine liver after intravenous injection of vectors expressing native or Tat-modified β-glucuronidase.
Figure 3: β-Glucuronidase activity in nonhepatic tissues after intravenous injection of mice with vectors expressing native or Tat-modified β-glucuronidase.
Figure 4: Distribution and activity of GFP and β-glucuronidase in brain.
Figure 5: Expression of β-glucuronidase or β-glucuronidase–Tat47–57 from transduced ependyma.

Similar content being viewed by others

References

  1. Meikle, P.J., Hopwood, J.J., Clague, A.E. & Carey, W.F. Prevalence of lysosomal storage disorders. JAMA 281, 249–254 (1999).

    Article  CAS  Google Scholar 

  2. O'Connor, L.H. et al. Enzyme replacement therapy for murine mucopolysaccharidosis type VII leads to improvements in behavior and auditory function. J. Clin. Invest. 101, 1394–1400 (1998).

    Article  CAS  Google Scholar 

  3. Frisella, W.A. et al. Intracranial injection of recombinant adeno-associated virus improves cognitive function in a murine model of mucopolysaccharidosis type VII. Mol. Ther. 3, 351–358 (2001).

    Article  CAS  Google Scholar 

  4. Schwarze, S.R., Ho, A., Vocero-Akbani, A. & Dowdy, S.F. In vivo protein transduction: delivery of a biologically active protein into the mouse. Science 285, 1569–1572 (1999).

    Article  CAS  Google Scholar 

  5. Lewin, M. et al. Tat peptide-derivatized magnetic nanoparticles allow in vivo tracking and recovery of progenitor cells. Nat. Biotechnol. 18, 410–414 (2000).

    Article  CAS  Google Scholar 

  6. Fawell, S. et al. Tat-mediated delivery of heterologous proteins into cells. Proc. Natl. Acad. Sci. USA 91, 664–668 (1994).

    Article  CAS  Google Scholar 

  7. Derossi, D. et al. Cell internalization of the third helix of the antennapedia homeodomain is receptor-independent. J. Biol. Chem. 271, 18188–18193 (1996).

    Article  CAS  Google Scholar 

  8. Ghodsi, A. et al. Extensive β-glucuronidase activity in murine CNS after adenovirus mediated gene transfer to brain. Hum. Gene Ther. 9, 2331–2340 (1998).

    Article  CAS  Google Scholar 

  9. Mann, D.A. & Frankel, A.D. Endocytosis and targeting of exogenous HIV-1 tat protein. EMBO J. 10, 1733–1739 (1991).

    Article  CAS  Google Scholar 

  10. Stein, C.S., Ghodsi, A., Derksen, T. & Davidson, B.L. Systemic and central nervous system correction of lysosomal storage in mucopolysaccharidosis type VII mice. J. Virol. 73, 3424–3429 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Ghodsi, A. et al. Systemic hyperosmolality improves β-glucuronidase distribution and pathology in murine MPS VII brain following intraventricular gene transfer. Exp. Neurol. 160, 109–116 (1999).

    Article  CAS  Google Scholar 

  12. Davidson, B.L. et al. Recombinant AAV type 2, 4 and 5 vectors: transduction of variant cell types and regions in the mammalian CNS. Proc. Natl. Acad. Sci. USA 97, 3428–3432 (2000).

    Article  CAS  Google Scholar 

  13. Anderson, R.D., Haskell, R.E., Xia, H., Roessler, B.J. & Davidson, B.L. A simple method for the rapid generation of recombinant adenovirus vectors. Gene Ther. 7, 1034–1038 (2000).

    Article  CAS  Google Scholar 

  14. Glaser, J.H. & Sly, W.S. Beta-glucuronidase deficiency mucopolysaccharidosis: methods for enzymatic diagnosis. J. Lab. Clin. Med. 82, 969–977 (1973).

    CAS  PubMed  Google Scholar 

  15. Sands, M.S. et al. Enzyme replacement therapy for murine mucopolysaccharidosis type VII. J. Clin. Invest. 93, 2324–2331 (1994).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Todd Derksen, Christine McLennan, Inês Martins, Christopher van de Wetering, Bridget Zimmerman, Paul Reimann, and Chad Stocker for assistance. This work was supported by the NIH (HD33531, NS34568, and DK54759), the American Heart Association (HX), the State of Iowa Biosciences Initiative (Q.M.), and the Roy J. Carver Trust (B.L.D.).

Author information

Author notes

  1. Haibin Xia and Qinwen Mao: These two authors contributed equally to this work.

Authors and Affiliations

  1. Department of Internal Medicine, Program in Gene Therapy, University of Iowa College of Medicine, Iowa City, 52242, IA, USA

    Haibin Xia, Qinwen Mao & Beverly L. Davidson

  2. Department of Neurology, Program in Gene Therapy, University of Iowa College of Medicine, Iowa City, 52242, IA, USA

    Beverly L. Davidson

  3. Department of Physiology and Biophysics, Program in Gene Therapy, University of Iowa College of Medicine, Iowa City, 52242, IA, USA

    Beverly L. Davidson

Authors
  1. Haibin Xia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qinwen Mao
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Beverly L. Davidson
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Beverly L. Davidson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Xia, H., Mao, Q. & Davidson, B. The HIV Tat protein transduction domain improves the biodistribution of β-glucuronidase expressed from recombinant viral vectors. Nat Biotechnol 19, 640–644 (2001). https://doi.org/10.1038/90242

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/90242

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing