title: Targeted protein destabilization reveals an estrogen-mediated ER stress response.
abstract: Accumulation of unfolded proteins within the endoplasmic reticulum (ER) of eukaryotic cells leads to an unfolded protein response (UPR) that either restores homeostasis or commits the cells to apoptosis. Tools traditionally used to study the UPR are proapoptotic and thus confound analysis of long-term cellular responses to ER stress. Here, we describe an ER-localized HaloTag (ERHT) protein that can be conditionally destabilized using a small-molecule hydrophobic tag (HyT36). Treatment of ERHT-expressing cells with HyT36 induces acute, resolvable ER stress that results in transient UPR activation without induction of apoptosis. Transcriptome analysis of late-stage responses to this UPR stimulus reveals a link between UPR activity and estrogen signaling.
authors.affiliation: 1] Department of Chemistry, Yale University New Haven, Connecticut, USA. [2].
authors.forename: Kanak
authors.initials: K
authors.lastname: Raina
authors.affiliation: 1] Department of Molecular, Cellular and Developmental Biology, Yale University New Haven, Connecticut, USA. [2].
authors.forename: Devin J
authors.initials: DJ
authors.lastname: Noblin
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University New Haven, Connecticut, USA.
authors.forename: Yevgeniy V
authors.initials: YV
authors.lastname: Serebrenik
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University New Haven, Connecticut, USA.
authors.forename: Alison
authors.initials: A
authors.lastname: Adams
authors.affiliation: Molecular Biophysics and Biochemistry, Yale University New Haven, Connecticut, USA.
authors.forename: Connie
authors.initials: C
authors.lastname: Zhao
authors.affiliation: 1] Department of Chemistry, Yale University New Haven, Connecticut, USA. [2] Department of Molecular, Cellular and Developmental Biology, Yale University New Haven, Connecticut, USA. [3] Department of Pharmacology, Yale University New Haven, Connecticut, USA.
authors.forename: Craig M
authors.identifier: 0000000284562005
authors.initials: CM
authors.lastname: Crews
mesh_terms: D000218:Adamantane / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D017209:Apoptosis; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D004967:Estrogens / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D055550:Protein Stability / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D019284:Thapsigargin / Q000494:pharmacology; D014415:Tunicamycin / Q000494:pharmacology; D056811:Unfolded Protein Response / Q000187:drug effects*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D004967:Estrogens; C000595187:HyT36 compound; D008565:Membrane Proteins; D014415:Tunicamycin; D019284:Thapsigargin; D000218:Adamantane