GenomeRNAi - a database for RNAi phenotypes and reagents
TitleScreen TitleAssayBiomodelSpecies
A functional RNAi screen links O-GlcNAc modification of ribosomal proteins to stress granule and processing body assembly. Ohn et al. (2008) Stress granule (SG) and processing body (PB) assembly G3BP1 and DCP1a protein expressionU2OS (RDG3)H. sapiens


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.

Screen details

Stable Id: GR00203-A
Screen title: Stress granule (SG) and processing body (PB) assembly
Assay: G3BP1 and DCP1a protein expression
Method: Fluorescence
Scope: Druggable genes
Screen type: Cell-based
Species: Homo sapiens
Biosource: Cell line
Biomodel: U2OS (RDG3)
Library: Dharmacon, SMARTpool
Reagent type: siRNA
Score type: Percentage of cells with SGs or PBs
Cutoff: Complex criteria