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Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration

Nature Medicine volume 4, pages 1449–1452 (1998)Cite this article

Cellular manipulation by transfection or viral introduction of cDNA expression vectors and microinjection of proteins into cells presents various difficulties, including massive overexpression, broad cell-to-cell intracellular concentration ranges of expressed proteins and low percentage of cells targeted1,2. Moreover, use of antisense approaches to manipulate intracellular processes have both specific gene and cell-type restrictions. Thus, the ability to manipulate cellular processes by the introduction of full-length proteins in a concentration-dependent fashion into 100% of cells would alleviate these technological problems.

In 1988, Green3 and Frankel4 independently discovered HIV TAT protein is able to cross cell membranes. In 1994, Fawell et al.5 demonstrated that chemically cross-linking a 36-amino acid domain of TAT to heterologous proteins conferred the ability to transduce into cells. Other transduction domains were identified that reside in the Antennapedia (Antp) protein from Drosophila6 and HSV VP22 protein from HSV (ref. 7). Although the exact mechanism of transduction across cellular membranes remains unclear, small Antp peptides have been shown to transduce into cells at 4 °C in a receptorless fashion8, indicating that all cell types potentially can be targeted by this method.

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Figure 1: The pTAT-HA vector and purification protocol.
Figure 2: Analysis of transduced proteins.
Figure 3: In vivo biochemical and biological properties of transduced proteins.
Figure 4: Induction of cell scattering and filopodia by TAT-p27wt protein.

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Acknowledgements

We thank J. Roberts for wild-type p27 cDNA, J. Massague for p27 kkpv mutant cDNA; E. Harlow for Cdk2-DN cDNA; M. Dustin for confocal microscopy; and all members of the Dowdy lab for critical input. This work was supported by an NIH-MSTP fellowship (E.L.S.) and the Howard Hughes Medical Institute (S.F.D.).

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  1. Howard Hughes Medical Institute and Division of Molecular Oncology, Depts of Pathology and Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA H.N. & A.M.V.-A. contributed equally

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  1. Steven F. Dowdy

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  1. Hikaru Nagahara
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Correspondence to Steven F. Dowdy.

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Nagahara, H., Vocero-Akbani, A., Snyder, E. et al. Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration. Nat Med 4, 1449–1452 (1998). https://doi.org/10.1038/4042

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