RESUMEN
The LIR motif-docking site (LDS) of Atg8/LC3 proteins is essential for the binding of LC3-interacting region (LIR)-containing proteins and their subsequent degradation by macroautophagy/autophagy. In our recent study, we created a mutated LDS site in Atg8a, the <i>Drosophila</i> homolog of Atg8/LC3 and found that LDS mutants accumulate known autophagy substrates and have reduced lifespan. We also conducted quantitative proteomics analyses and identified several proteins that are enriched in the LDS mutants, including Gmap (Golgi microtubule-associated protein). Gmap contains a LIR motif and accumulates in LDS mutants. We showed that Gmap and Atg8a interact in a LIR-LDS dependent manner and that the Golgi size and morphology are altered in Atg8a-LDS and Gmap-LIR motif mutants. Our findings highlight a role for Gmap in the regulation of Golgiphagy.
Asunto(s)
Autofagia , Macroautofagia , Secuencias de Aminoácidos , Animales , Familia de las Proteínas 8 Relacionadas con la Autofagia/metabolismo , Drosophila/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Control de CalidadRESUMEN
Selective autophagy receptors and adapters contain short linear motifs called LIR motifs (LC3-interacting region), which are required for the interaction with the Atg8-family proteins. LIR motifs bind to the hydrophobic pockets of the LIR motif docking site (LDS) of the respective Atg8-family proteins. The physiological significance of LDS docking sites has not been clarified in vivo. Here, we show that Atg8a-LDS mutant Drosophila flies accumulate autophagy substrates and have reduced lifespan. Using quantitative proteomics to identify the proteins that accumulate in Atg8a-LDS mutants, we identify the cis-Golgi protein GMAP (Golgi microtubule-associated protein) as a LIR motif-containing protein that interacts with Atg8a. GMAP LIR mutant flies exhibit accumulation of Golgi markers and elongated Golgi morphology. Our data suggest that GMAP mediates the turnover of Golgi by selective autophagy to regulate its morphology and size via its LIR motif-mediated interaction with Atg8a.
Asunto(s)
Drosophila , Proteínas Asociadas a Microtúbulos , Secuencias de Aminoácidos , Animales , Autofagia , Familia de las Proteínas 8 Relacionadas con la Autofagia/genética , Drosophila/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismoRESUMEN
Hunger drives food-seeking behaviour and controls adaptation of organisms to nutrient availability and energy stores. Lipids constitute an essential source of energy in the cell that can be mobilised during fasting by autophagy. Selective degradation of proteins by autophagy is made possible essentially by the presence of LIR and KFERQ-like motifs. Using in silico screening of Drosophila proteins that contain KFERQ-like motifs, we identified and characterized the adaptor protein Arouser, which functions to regulate fat storage and mobilisation and is essential during periods of food deprivation. We show that hypomorphic arouser mutants are not satiated, are more sensitive to food deprivation, and are more aggressive, suggesting an essential role for Arouser in the coordination of metabolism and food-related behaviour. Our analysis shows that Arouser functions in the fat body through nutrient-related signalling pathways and is degraded by endosomal microautophagy. Arouser degradation occurs during feeding conditions, whereas its stabilisation during non-feeding periods is essential for resistance to starvation and survival. In summary, our data describe a novel role for endosomal microautophagy in energy homeostasis, by the degradation of the signalling regulatory protein Arouser.
Asunto(s)
Adaptación Fisiológica , Drosophila/fisiología , Endosomas/metabolismo , Microautofagia , Inanición , Animales , Cromatografía Liquida , Proteínas de Drosophila/química , Proteínas de Drosophila/metabolismo , Insulina/metabolismo , Metabolismo de los Lípidos , Nutrientes/metabolismo , Proteoma , Proteómica/métodos , Serina-Treonina Quinasas TOR/metabolismo , Espectrometría de Masas en TándemRESUMEN
Autophagy is a highly conserved catabolic process in which cytoplasmic material is recycled under various conditions of cellular stress, preventing cell damage and promoting survival in the event of energy or nutrient shortage, or in response to various cytotoxic insults. Autophagy is also responsible for the removal of aggregated proteins and damaged organelles, playing a vital role in the quality control of proteins and organelles. Impairment of autophagy has been linked to various diseases, including cancer and neurodegenerative disorders, making it a very interesting process for further research. Recent research highlighted that autophagy is not random and can be selective, making it even more important to understand the molecular mechanisms of selectivity at the organismal level. Drosophila has been demonstrated to be an excellent animal model for studying selective autophagy, as the autophagic machinery is highly conserved, although much is still left to be explored. In this review, an overview of autophagy and its selectivity in Drosophila will be presented.
Asunto(s)
Proteínas Relacionadas con la Autofagia/metabolismo , Autofagia , Drosophila/citología , Drosophila/metabolismo , AnimalesRESUMEN
BACKGROUND: Genomic sequencing technologies have made the possibility of population screening for whole panels of genetic disorders more feasible than ever before. As one of the most common single gene disorders affecting the UK population, hemophilia is an attractive candidate to include on such screening panels. However, very little is known about views toward genetic screening amongst people with hemophilia or their family members, despite the potential for a wide range of impacts on them. METHODS: Twenty-two in-depth qualitative interviews were undertaken to explore the views of adults with hemophilia and their family members, recruited through the Haemophilia Society UK. These interviews were used to develop a survey, the Haemophilia Screening Survey (UK), which was distributed in paper and online format through the support group, receiving 327 returns between January and June 2018. RESULTS: Fifty-seven per cent of the sample supported preconception carrier screening of the population for hemophilia, and 59% supported prenatal carrier screening. Key reasons for support included a desire to reduce pregnancy terminations and increase awareness of hemophilia. Despite support for screening however, 90% of the sample disagreed with pregnancy terminations for hemophilia. CONCLUSIONS: Families and adults living with hemophilia are more supportive of screening for information and preparation purposes than to prevent boys with hemophilia from being born. A distinction was made between preventing the disease and preventing the lives of people with it, with support shown for the use of screening to achieve the former, but not at the expense of the latter.