A novel role for the Drosophila epsin (lqf): involvement in autophagy.
Screening P-element-induced mutant collections, 52 lines were selected as potentially defected ones in endocytosis or autophagy. After excluding those which were rescued by 20-hydroxyecdysone treatment, the exact position of the inserted P-element was determined in the remaining lines. In the case of l(3)S011027 stock, the liquid facets (lqf) gene was affected which codes an epsin-homolog protein in Drosophila. We reveal that Lqf is essential to the receptor-mediated endocytosis of larval serum proteins (LSPs) in the larval fat body cells of Drosophila. In l(3)S011027 line, lack of Lqf fails the formation of autophagosomes thus leading to the arrest of destroying of trophocytes. Transgenic larvae carrying Lqf-RNAi construct were unable to generate endocytic and autophagic vacuoles and led to a prolonged larval stage. On the other hand, Lqf protein showed an exclusive colocalization with the LysoTracker Red- or GFP-Atg8a labeled autophagosomes. By using the antiserum generated against the fifth exon of lqf, we demonstrated that prior to the onset of developmental autophagy the Lqf protein was present in the nucleus of fat body cell, but thereafter the protein was localized in the territory of endocytic and autophagic vacuoles. The fact that the inhibition of the target of rapamycin (TOR) did not restore the autophagic process and the normal development in the case of lqf mutant larvae points to that the Lqf is downstream to the TOR, the central kinase of the autophagy pathway.
Csikós
György
G
Department of Anatomy, Cell and Developmental Biology, Eötvös Lóránd University, Budapest, Hungary.
Lippai
Mónika
M
Lukácsovich
Tamás
T
Juhász
Gábor
G
Henn
László
L
Erdélyi
Miklós
M
Maróy
Péter
P
Sass
Miklós
M
D000165:Acridine Orange / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism*; D000483:Alleles; D000588:Amines / Q000378:metabolism; D000818:Animals; D001343:Autophagy / Q000235:genetics; D002999:Clone Cells; D004251:DNA Transposable Elements / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D004441:Ecdysterone / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects; D005216:Fat Body / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D017344:Genes, Insect; D005816:Genetic Complementation Test; D007106:Immune Sera; D007814:Larva / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D008938:Mitosis / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D010588:Phagosomes / Q000187:drug effects / Q000648:ultrastructure; D034622:RNA Interference / Q000187:drug effects; D020123:Sirolimus / Q000494:pharmacology; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism*
D033942:Adaptor Proteins, Vesicular Transport; D000588:Amines; D004251:DNA Transposable Elements; D029721:Drosophila Proteins; D007106:Immune Sera; C494336:Lqf protein, Drosophila; C493330:Red DND-99; D033921:Vesicular Transport Proteins; C114154:epsin; D004441:Ecdysterone; D000165:Acridine Orange; D020123:Sirolimus