title: A functional RNAi screen links O-GlcNAc modification of ribosomal proteins to stress granule and processing body assembly.
abstract: Stress granules (SGs) and processing bodies (PBs) are microscopically visible ribonucleoprotein granules that cooperatively regulate the translation and decay of messenger RNA. Using an RNA-mediated interference-based screen, we identify 101 human genes required for SG assembly, 39 genes required for PB assembly, and 31 genes required for coordinate SG and PB assembly. Although 51 genes encode proteins involved in mRNA translation, splicing and transcription, most are not obviously associated with RNA metabolism. We find that several components of the hexosamine biosynthetic pathway, which reversibly modifies proteins with O-linked N-acetylglucosamine (O-GlcNAc) in response to stress, are required for SG and PB assembly. O-GlcNAc-modified proteins are prominent components of SGs but not PBs, and include RACK1 (receptor for activated C kinase 1), prohibitin-2, glyceraldehyde-3-phosphate dehydrogenase and numerous ribosomal proteins. Our results suggest that O-GlcNAc modification of the translational machinery is required for aggregation of untranslated messenger ribonucleoproteins into SGs. The lack of enzymes of the hexosamine biosynthetic pathway in budding yeast may contribute to differences between mammalian SGs and related yeast EGP (eIF4E, 4G and Pab1 containing) bodies.
authors.affiliation: Division of Rheumatology, Immunology and Allergy, Brigham and Women's Hospital, Harvard Medical School, 1 Jimmy Fund Way, Boston, Massachusetts 02115, USA.
authors.forename: Takbum
authors.initials: T
authors.lastname: Ohn
authors.forename: Nancy
authors.initials: N
authors.lastname: Kedersha
authors.forename: Tyler
authors.initials: T
authors.lastname: Hickman
authors.forename: Sarah
authors.initials: S
authors.lastname: Tisdale
authors.forename: Paul
authors.initials: P
authors.lastname: Anderson
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism*; D000818:Animals; D018053:Arsenites / Q000494:pharmacology; D002460:Cell Line; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000378:metabolism*; D022083:Cytoplasmic Structures / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D034622:RNA Interference / Q000187:drug effects; D012261:Ribonucleoproteins / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D018053:Arsenites; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins; C015001:arsenite; D000117:Acetylglucosamine
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