Rapid regulation of protein activity in fission yeast
- Equal contributors
1 Department of Cell Biology, Rikshospitalet-Radiumhospitalet Medical Centre, Montebello, 0310 Oslo, Norway
2 Department of Zoology, University of Oxford, South Parks Road, Oxford OX1 3PS, UK
3 Department of Molecular Biology, University of Copenhagen, Københavns Biocenter, Ole Maaløes Vej 5, 2200 Copenhagen N, Denmark
4 Samuel Lunenfeld Research Institute, Room 1080, Mount Sinai Hospital, 600 University Avenue, Toronto Ontario, M5G 1X5, Canada
BMC Cell Biology 2008, 9:23 doi:10.1186/1471-2121-9-23Published: 5 May 2008
The fission yeast Schizosaccharomyces pombe is widely-used as a model organism for the study of a broad range of eukaryotic cellular processes such as cell cycle, genome stability and cell morphology. Despite the availability of extensive set of genetic, molecular biological, biochemical and cell biological tools for analysis of protein function in fission yeast, studies are often hampered by the lack of an effective method allowing for the rapid regulation of protein level or protein activity.
In order to be able to regulate protein function, we have made use of a previous finding that the hormone binding domain of steroid receptors can be used as a regulatory cassette to subject the activity of heterologous proteins to hormonal regulation. The approach is based on fusing the protein of interest to the hormone binding domain (HBD) of the estrogen receptor (ER). The HBD tag will attract the Hsp90 complex, which can render the fusion protein inactive. Upon addition of estradiol the protein is quickly released from the Hsp90 complex and thereby activated. We have tagged and characterised the induction of activity of four different HBD-tagged proteins. Here we show that the tag provided the means to effectively regulate the activity of two of these proteins.
The estradiol-regulatable hormone binding domain provides a means to regulate the function of some, though not all, fission yeast proteins. This system may result in very quick and reversible activation of the protein of interest. Therefore it will be a powerful tool and it will open experimental approaches in fission yeast that have previously not been possible. Since fission yeast is a widely-used model organism, this will be valuable in many areas of research.