The anchor-away technique: rapid, conditional establishment of yeast mutant phenotypes

Article Article scientifique Article The anchor-away technique: rapid, conditional establishment of yeast mutant phenotypes eng 2008-09-26 author Haruki Hirohito 650862 author Nishikawa Junichi 629865 author Laemmli Ulrich Karl 25441 925-932 10.1016/j.molcel.2008.07.020 18922474 1097-2765 The anchor-away (AA) technique depletes the nucleus of Saccharomyces cerevisiae of a protein of interest (the target) by conditional tethering to an abundant cytoplasmic protein (the anchor) by appropriate gene tagging and rapamycin-dependent heterodimerization. Taking advantage of the massive flow of ribosomal proteins through the nucleus during maturation, a protein of the large subunit was chosen as the anchor. Addition of rapamycin, due to formation of the ternary complex, composed of the anchor, rapamycin, and the target, then results in the rapid depletion of the target from the nucleus. All 43 tested genes displayed on rapamycin plates the expected defective growth phenotype. In addition, when examined functionally, specific mutant phenotypes were obtained within minutes. These are genes involved in protein import, RNA export, transcription, sister chromatid cohesion, and gene silencing. The AA technique is a powerful tool for nuclear biology to dissect the function of individual or gene pairs in synthetic, lethal situations. Cell Nucleus / drug effects Cell Nucleus / genetics Cell Proliferation / drug effects Genes, Fungal Genetic Complementation Test Genetic Techniques Mutagenesis / drug effects Phenotype Ribosomes / drug effects Ribosomes / metabolism Saccharomyces cerevisiae / cytology Saccharomyces cerevisiae / drug effects Saccharomyces cerevisiae / genetics Saccharomyces cerevisiae Proteins / genetics Saccharomyces cerevisiae Proteins / metabolism Sirolimus / pharmacology Time Factors Transcription, Genetic / drug effects Dewey 540 39 Département de biochimie 144 false Molecular cell 31 6 9bcc6184-2e6a-47b0-993d-ab6e5a6df1c8 Article (Published version) Public Article-scientifique-from-unpaywall.pdf 956951 application/pdf CC BY-4.0 openAccess Faculté des sciences / Section de chimie et biochimie / Département de biochimie

Title	Published in	Access level	OA Policy	Year	Views	Downloads

The anchor-away technique: rapid, conditional establishment of yeast mutant phenotypes	Molecular cell			2008	219	377

Archive ouverte UNIGE

Haruki, Hirohito