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Activity of the yeast Tat2p tryptophan permease is sensitive to the anti-tumor agent 4-phenylbutyrate

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Abstract

4-Phenylbutyrate (PB) induces differentiation and is being intensively studied as a treatment for brain, prostate, breast, and hematopoietic cancer. While many different primary targets for PB have been proposed, the mechanism by which it causes cellular differentiation remains unknown. To identify the primary cellular target, we investigated its effects on Saccharomyces cerevisiae and showed that it inhibits tryptophan transport. We show here that PB and sorbic acid induce an ubiquitin-dependent turnover of the tryptophan permease Tat2p. However, the inhibition of transport is not a consequence of the loss of Tat2p, since it also occurs when turnover is prevented by deleting the Tat2p ubiquitination sites. When we tested the effects of PB and other growth inhibitory agents on the growth of amino acid auxotrophs, we found that several auxotrophs are hypersensitive to a number of chemically unrelated agents, including PB and some, but not all, weak acids; and this sensitivity is due to the inhibition of amino acid transport. For the inhibitory weak acids, inhibition is not confined to aromatic amino acid auxotrophs, nor is it a general weak acid stress response, since the degree of inhibition is independent of weak acid hydrophobicity and pKa. Our results show that diverse agents affect the activity of the Tat2p permease rather than its stability and suggest the hypothesis that the anti-neoplastic action of PB is due to a decrease in the activity of surface receptors or other membrane proteins needed to maintain the transformed state.

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Acknowledgements

We wish to thank the following individuals for generously supplying plasmids and strains: Bruno Andre (Universite Libre de Bruxelles, Belgium), Morton C. Kielland-Brandt (Carlsberg Laboratory, Copenhagen, Denmark), Frank Boschelli (Wyeth Research, Pearl River, N.Y., USA), Andrew Grzanowski (School of Veterinary Medicine, Michigan State University, Lansing, Mich., USA), Michael N. Hall (University of Basel, Basel, Switzerland), Randal J. Kaufman and Martin Schröder (University of Michigan Medical Center, Ann Arbor, Mich., USA), and Per O. Ljungdahl (Ludwig Institute for Cancer Research in Stockholm, Sweden). We also thank Nick and Amy Davis (WSU School of Medicine, Detroit, Mich., USA) for advice on the Western transfer analysis.

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  1. Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, 540 E. Canfield Ave, Detroit, MI 48201, USA

    Ming Liu, William S. A. Brusilow & Richard Needleman

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  1. Ming Liu
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  3. Richard Needleman
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Correspondence to Richard Needleman.

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Liu, M., Brusilow, W.S.A. & Needleman, R. Activity of the yeast Tat2p tryptophan permease is sensitive to the anti-tumor agent 4-phenylbutyrate. Curr Genet 46, 256–268 (2004). https://doi.org/10.1007/s00294-004-0531-7

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