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Rapid regulation of nuclear proteins by rapamycin-induced translocation in fission yeast.

ABSTRACT: 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.

SUBMITTER: Ding L

PROVIDER: S-EPMC4106978 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature

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Rapid regulation of nuclear proteins by rapamycin-induced translocation in fission yeast.

Ding Lin L Laor Dana D Weisman Ronit R Forsburg Susan L SL

Yeast (Chichester, England) 20140429 7

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 ...[more]

PMID: 24733494

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