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Deciphering the function of UbcD4 in selective autophagy in Drosophila melanogaster
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Prisco, Laura (2022) Deciphering the function of UbcD4 in selective autophagy in Drosophila melanogaster. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b3929619
Abstract
Autophagy is a lysosomal degradative pathway, which recycles cytoplasmic contents. Proteins encoded by autophagy related genes (ATGs) regulate autophagy. Among these, Atg8/LC3 proteins are essential for formation of autophagosomes. Atg8 homologue interactions are mediated by LIR (LC3-interacting region) motifs. Ubiquitination plays an important role in targeting specific proteins for degradation by autophagosomes. Several receptors involved in this process contain an interaction domain for ubiquitin moieties.
Screening for LIR motifs among proteins containing a ubiquitin-associated (UBA) domain in Drosophila melanogaster, revealed the ubiquitin conjugating enzyme UbcD4. UbcD4 was found to aggregate in Atg8a mutant (autophagy deficient) fly brains and colocalize with ubiquitinated protein aggregates and ref(2)P (refractory to sigma P, a protein required for ubiquitinated protein aggregate formation).
The project intends to understand the function of UbcD4 in selective autophagy in Drosophila. GST pulldown and immunoprecipitation of UbcD4 and UbcD4 LIR mutant with Atg8a found that the two proteins interact regardless of the LIR motif. Further proteomic analysis revealed the wider UbcD4 structure to be implicated in this interaction, namely a glycine-rich linker region and a disordered region located in the C-terminal domain (CTD) containing a methionine residue that is oxidised only in the UbcD4 wild type variant. Furthermore, the creation of a UbcD4 CRISPR mutant showed UbcD4 has an effect on lifespan (with gender bias), climbing ability and the formation of ref(2)P and ubiquitinated protein aggrregates in Drosophila guts. The results obtainted suggest UbcD4 has a regulatory role in selective autophagy via interaction with Atg8a and affects the formation of ubiquitinated protein aggregates despite not impacting ubiquitinated protein expression.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QH Natural history Q Science > QH Natural history > QH301 Biology Q Science > QP Physiology |
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Library of Congress Subject Headings (LCSH): | Autophagic vacuoles, Drosophila melanogaster, Ubiquitin, Protein-protein interactions | ||||
Official Date: | October 2022 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Nezis, Ioannis P. | ||||
Sponsors: | Biotechnology and Biological Sciences Research Council (Great Britain) ; Midlands Integrative Biosciences Training Partnership | ||||
Format of File: | |||||
Extent: | 166 pages : colour illustrations | ||||
Language: | eng |
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