title: Disease-related amyloidogenic variants of human lysozyme trigger the unfolded protein response and disturb eye development in Drosophila melanogaster.
abstract: We have created a Drosophila model of lysozyme amyloidosis to investigate the in vivo behavior of disease-associated variants. To achieve this objective, wild-type (WT) protein and the amyloidogenic variants F57I and D67H were expressed in Drosophila melanogaster using the UAS-gal4 system and both the ubiquitous and retinal expression drivers Act5C-gal4 and gmr-gal4. The nontransgenic w(1118) Drosophila line was used as a control throughout. We utilized ELISA experiments to probe lysozyme protein levels, scanning electron microscopy for eye phenotype classification, and immunohistochemistry to detect the unfolded protein response (UPR) activation. We observed that expressing the destabilized F57I and D67H lysozymes triggers UPR activation, resulting in degradation of these variants, whereas the WT lysozyme is secreted into the fly hemolymph. Indeed, the level of WT was up to 17 times more abundant than the variant proteins. In addition, the F57I variant gave rise to a significant disruption of the eye development, and this correlated to pronounced UPR activation. These results support the concept that the onset of familial amyloid disease is linked to an inability of the UPR to degrade completely the amyloidogenic lysozymes prior to secretion, resulting in secretion of these destabilized variants, thereby leading to deposition and associated organ damage.
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge, UK.
authors.forename: Janet R
authors.initials: JR
authors.lastname: Kumita
authors.forename: Linda
authors.initials: L
authors.lastname: Helmfors
authors.forename: Jocy
authors.initials: J
authors.lastname: Williams
authors.forename: Leila M
authors.initials: LM
authors.lastname: Luheshi
authors.forename: Linda
authors.initials: L
authors.lastname: Menzer
authors.forename: Mireille
authors.initials: M
authors.lastname: Dumoulin
authors.forename: David A
authors.initials: DA
authors.lastname: Lomas
authors.forename: Damian C
authors.initials: DC
authors.lastname: Crowther
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Dobson
authors.forename: Ann-Christin
authors.initials: AC
authors.lastname: Brorsson
mesh_terms: D000686:Amyloidosis / Q000201:enzymology* / Q000473:pathology; D000818:Animals; D030801:Animals, Genetically Modified; D004268:DNA-Binding Proteins / Q000235:genetics; D004195:Disease Models, Animal; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster; D059865:Endoplasmic Reticulum Stress / Q000502:physiology; D005124:Eye Abnormalities / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics; D006458:Hemolymph / Q000201:enzymology; D006801:Humans; D008297:Male; D008675:Metamorphosis, Biological / Q000502:physiology; D008855:Microscopy, Electron, Scanning; D009113:Muramidase / Q000235:genetics / Q000378:metabolism*; D017956:Photoreceptor Cells, Invertebrate / Q000201:enzymology / Q000473:pathology / Q000648:ultrastructure; D012995:Solubility; D056811:Unfolded Protein Response / Q000502:physiology*
chemical_list: D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; C517798:Xbp1 protein, Drosophila; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D009113:Muramidase