title: A light-inducible organelle-targeting system for dynamically activating and inactivating signaling in budding yeast.
abstract: Protein localization plays a central role in cell biology. Although powerful tools exist to assay the spatial and temporal dynamics of proteins in living cells, our ability to control these dynamics has been much more limited. We previously used the phytochrome B- phytochrome-interacting factor light-gated dimerization system to recruit proteins to the plasma membrane, enabling us to control the activation of intracellular signals in mammalian cells. Here we extend this approach to achieve rapid, reversible, and titratable control of protein localization for eight different organelles/positions in budding yeast. By tagging genes at the endogenous locus, we can recruit proteins to or away from their normal sites of action. This system provides a general strategy for dynamically activating or inactivating proteins of interest by controlling their localization and therefore their availability to binding partners and substrates, as we demonstrate for galactose signaling. More importantly, the temporal and spatial precision of the system make it possible to identify when and where a given protein's activity is necessary for function, as we demonstrate for the mitotic cyclin Clb2 in nuclear fission and spindle stabilization. Our light-inducible organelle-targeting system represents a powerful approach for achieving a better understanding of complex biological systems.
authors.affiliation: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA 94158, USA.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Yang
authors.forename: Anna Payne-Tobin
authors.initials: AP
authors.lastname: Jost
authors.forename: Orion D
authors.initials: OD
authors.lastname: Weiner
authors.forename: Chao
authors.initials: C
authors.lastname: Tang
mesh_terms: D016203:CDC2 Protein Kinase / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D019926:Cyclin B / Q000235:genetics / Q000378:metabolism; D008027:Light; D009680:Nuclear Fission; D062308:Optogenetics; D015388:Organelles / Q000378:metabolism; D051939:Phytochrome B / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D055550:Protein Stability; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D008941:Spindle Apparatus / Q000378:metabolism; D059008:Time-Lapse Imaging
chemical_list: C093749:CLB2 protein, S cerevisiae; D019926:Cyclin B; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D051939:Phytochrome B; D016203:CDC2 Protein Kinase