Premium
Rapid regulation of nuclear proteins by rapamycin‐induced translocation in fission yeast
Author(s) -
Ding Lin,
Laor Dana,
Weisman Ronit,
Forsburg Susan L.
Publication year - 2014
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.3014
Subject(s) - schizosaccharomyces pombe , biology , yeast , cytoplasm , nuclear protein , nuclear transport , fission , microbiology and biotechnology , schizosaccharomyces , function (biology) , budding yeast , gene , saccharomyces cerevisiae , genetics , computational biology , cell nucleus , transcription factor , physics , quantum mechanics , neutron
Genetic analysis of protein function requires a rapid means of inactivating the gene under study. Typically, this exploits temperature‐sensitive mutations or promoter shut‐off techniques. We report the adaptation to Schizosaccharomyces pombe of the anchor‐away technique, originally designed in budding yeast by Laemmli lab. This method relies on a rapamycin‐mediated interaction between the FRB‐ and FKBP12‐binding domains to relocalize nuclear proteins of interest to the cytoplasm. We demonstrate a rapid nuclear depletion of abundant proteins as proof of principle. Copyright © 2014 John Wiley & Sons, Ltd.