Intranuclear protein transduction through a nucleoside salvage pathway

James E. Hansen, Chung Ming Tse, Grace Chan, Emil R. Heinze, Robert N. Nishimura, Richard H. Weisbart

Research output: Contribution to journalArticle

Abstract

Regulation of gene expression by intranuclear transduction of macromolecules such as transcription factors is an alternative to gene therapy for the treatment of numerous diseases. The identification of an effective intranuclear delivery vehicle and pathway for the transport of therapeutic macromolecules across plasma and nuclear membranes, however, has posed a significant challenge. The anti-DNA antibody fragment 3E10 Fv has received attention as a novel molecular delivery vehicle due to its penetration into living cells with specific nuclear localization, absence of toxicity, and successful delivery of therapeutic cargo proteins in vitro and in vivo. Elucidation of the pathway that allows 3E10 Fv to cross cell membranes is critical to the development of new molecular therapies. Here we show that 3E10 Fv penetrates cells through a nucleoside salvage transporter. 3E10 Fv is unable to penetrate into cells deficient in the equilibrative nucleoside transporter ENT2, and reconstitution of ENT2 into ENT2-deficient cells restores 3E10 Fv transport into cell nuclei. Our results represent the first demonstration of protein transport through a nucleoside salvage pathway. We expect that our finding will facilitate a variety of methods of gene regulation in the treatment of human diseases, open up new avenues of research in nucleoside salvage pathways, and enhance our understanding of the pathophysiology of autoimmune diseases.

Original languageEnglish (US)
Pages (from-to)20790-20793
Number of pages4
JournalJournal of Biological Chemistry
Volume282
Issue number29
DOIs
StatePublished - Jul 20 2007

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Hansen, J. E., Tse, C. M., Chan, G., Heinze, E. R., Nishimura, R. N., & Weisbart, R. H. (2007). Intranuclear protein transduction through a nucleoside salvage pathway. Journal of Biological Chemistry, 282(29), 20790-20793. https://doi.org/10.1074/jbc.C700090200