Department of Biochemistry and Molecular Pharmacology
Cell Biology | Fungi | Genetic Phenomena | Hormones, Hormone Substitutes, and Hormone Antagonists
The fission yeast Schizosaccharomyces pombe lacks a diverse toolkit of inducible promoters for experimental manipulation. Available inducible promoters suffer from slow induction kinetics, limited control of expression levels and/or a requirement for defined growth medium. In particular, no S. pombe inducible promoter systems exhibit a linear dose response, which would allow expression to be tuned to specific levels. We have adapted a fast, orthogonal promoter system with a large dynamic range and a linear dose response, based on β-estradiol-regulated function of the human estrogen receptor, for use in S. pombe. We show that this promoter system, termed Z3EV, turns on quickly, can reach a maximal induction of 20 fold, and exhibits a linear dose response over its entire induction range, with few off target effects. We demonstrate the utility of this system by regulating the mitotic inhibitor Wee1 to create a strain in which cell size is regulated by β-estradiol concentration. This promoter system will be of great utility for experimentally regulating gene expression in fission yeast.
cell biology, Schizosaccharomyces pombe, gene expression, estradiol
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The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.
DOI of Published Version
bioRxiv 092338; doi: https://doi.org/10.1101/092338. Link to preprint on bioRxiv service.
Now published in Yeast doi: 10.1002/yea.3235.
Ohira, Makoto J.; Hendrickson, David G.; McIsaac, R. Scott; and Rhind, Nicholas R., "An Estradiol-Inducible Promoter Enables Fast, Graduated Control of Gene Expression in Fission Yeast" (2017). University of Massachusetts Medical School Faculty Publications. 1554.
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