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Conditional Nuclear Import and Export of Yeast Proteins Using a Chemical Inducer of Dimerization

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Abstract

In eukaryotes, reversible shuttling between the nucleus and cytoplasm is an important regulatory mechanism, particularly for many kinases and transcription factors. Inspired by the natural system, we recently developed a technology to control protein position in budding yeast using a chemical inducer of dimerization (CID). In this method, a nuclear export or localization signal is reversibly appended to a protein of interest by the CID, which effectively places its subcellular location under direct control of the chemical stimulus. Here, we explicitly tested the ability of this system to direct the nucleocytoplasmic transport of a panel of 16 representative kinases and transcription factors. From this set, we found that 12 targets (75%) are susceptible to re-positioning, suggesting that this method might be applicable to a range of targets. Interestingly, the four proteins that resisted mislocalization (Fun20p, Hcm1p, Pho4p, and Ste12p) are known to engage in a large number of protein–protein contacts. We suspect that, for these highly connected targets, the strength of the chemical signal may be insufficient to drive mislocalization and that proteins with relatively few partners might be most amenable to this approach. Collectively, these studies provide a necessary framework for the design of large-scale applications.

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Acknowledgments

The authors thank C. Trevor, R. Lahey, and T. Cory for helpful comments and eager assistance. This work was supported by grants RSG-06-179-01-MBC from the American Cancer Society, DBI 0543017 from the National Science Foundation, and Basil O’Connor Award 5-FY05-1224 from the March of Dimes (to A.K.).

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Authors and Affiliations

  1. Department of Pathology, University of Michigan, Ann Arbor, MI, 48109-2216, USA

    Srikanth Patury & Jason E. Gestwicki

  2. The Life Sciences Institute, University of Michigan, 210 Washtenaw Ave, Ann Arbor, MI, 48109-2216, USA

    Srikanth Patury, Prasanthi Geda, Craig J. Dobry, Anuj Kumar & Jason E. Gestwicki

  3. Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI, 48109-2216, USA

    Prasanthi Geda, Craig J. Dobry & Anuj Kumar

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  1. Srikanth Patury
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  2. Prasanthi Geda
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  5. Jason E. Gestwicki
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Correspondence to Jason E. Gestwicki.

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Patury, S., Geda, P., Dobry, C.J. et al. Conditional Nuclear Import and Export of Yeast Proteins Using a Chemical Inducer of Dimerization. Cell Biochem Biophys 53, 127–134 (2009). https://doi.org/10.1007/s12013-009-9044-9

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