title: Nucleolar stress in Drosophila melanogaster: RNAi-mediated depletion of Nopp140.
abstract: Nucleolar stress results when ribosome biogenesis is disrupted. An excellent example is the human Treacher Collins syndrome in which the loss of the nucleolar chaperone, Treacle, leads to p53-dependent apoptosis in embryonic neural crest cells and ultimately to craniofacial birth defects. Here, we show that depletion of the related nucleolar phosphoprotein, Nopp140, in Drosophila melanogaster led to nucleolar stress and eventual lethality when multiple tissues were depleted of Nopp140. We used TEM, immuno-blot analysis and metabolic protein labeling to show the loss of ribosomes. Targeted loss of Nopp140 in larval wing discs caused Caspase-dependent apoptosis which eventually led to defects in the adult wings. These defects were not rescued by a p53 gene deletion, as the craniofacial defects were in the murine model of TCS, thus suggesting that apoptosis caused by nucleolar stress in Drosophila is induced by a p53-independent mechanism. Loss of Nopp140 in larval polyploid midgut cells induced premature autophagy as marked by the accumulation of mCherry-ATG8a into autophagic vesicles. We also found elevated phenoloxidase A3 levels in whole larval lysates and within the hemolymph of Nopp140-depleted larvae vs. hemolymph from parental genotype larvae. Phenoloxidase A3 enrichment was coincident with the appearance of melanotic tumors in the Nopp140-depleted larvae. The occurrence of apoptosis, autophagy and phenoloxidase A3 release to the hemolymph upon nucleolar stress correlated well with the demonstrated activation of Jun N-terminal kinase (JNK) in Nopp140-depleted larvae. We propose that JNK is a central stress response effector that is activated by nucleolar stress in Drosophila larvae.
authors: Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, USA.
authors: Allison
authors: A
authors: James
authors: Renford
authors: R
authors: Cindass
authors: Dana
authors: D
authors: Mayer
authors: Stephanie
authors: S
authors: Terhoeve
authors: Courtney
authors: C
authors: Mumphrey
authors: Patrick
authors: P
authors: DiMario
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002466:Cell Nucleolus / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D006458:Hemolymph / Q000378:metabolism; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics; D008342:Mandibulofacial Dysostosis / Q000235:genetics*; D014442:Monophenol Monooxygenase / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D016601:RNA-Binding Proteins; D013312:Stress, Physiological / Q000235:genetics
chemical_list: D029721:Drosophila Proteins; C453465:Nopp140 protein, Drosophila; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D014442:Monophenol Monooxygenase; D048031:JNK Mitogen-Activated Protein Kinases
keywords: Drosophila; Nopp140; apoptosis; autophagy; nucleolus