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| Item Type: | Article |
|---|---|
| Title: | Proteomic analysis of the ubiquitin landscape in the drosophila embryonic nervous system and the adult photoreceptor cells |
| Creators Name: | Ramirez, J. and Martinez, A. and Lectez, B. and Lee, S.Y. and Franco, M. and Barrio, R. and Dittmar, G. and Mayor, U. |
| Abstract: | Background: Ubiquitination is known to regulate physiological neuronal functions as well as to be involved in a number of neuronal diseases. Several ubiquitin proteomic approaches have been developed during the last decade but, as they have been mostly applied to non-neuronal cell culture, very little is yet known about neuronal ubiquitination pathways in vivo. Methodology/Principal Findings: Using an in vivo biotinylation strategy we have isolated and identified the ubiquitinated proteome in neurons both for the developing embryonic brain and for the adult eye of Drosophila melanogaster. Bioinformatic comparison of both datasets indicates a significant difference on the ubiquitin substrates, which logically correlates with the processes that are most active at each of the developmental stages. Detection within the isolated material of two ubiquitin E3 ligases, Parkin and Ube3a, indicates their ubiquitinating activity on the studied tissues. Further identification of the proteins that do accumulate upon interference with the proteasomal degradative pathway provides an indication of the proteins that are targeted for clearance in neurons. Last, we report the proof-of-principle validation of two lysine residues required for nSyb ubiquitination. Conclusions/Significance: These data cast light on the differential and common ubiquitination pathways between the embryonic and adult neurons, and hence will contribute to the understanding of the mechanisms by which neuronal function is regulated. The in vivo biotinylation methodology described here complements other approaches for ubiquitome study and offers unique advantages, and is poised to provide further insight into disease mechanisms related to the ubiquitin proteasome system. |
| Keywords: | Amino Acid Sequence, Biotinylation, Cell Line, Drosophila Proteins, Green Fluorescent Proteins, Invertebrate Photoreceptor Cells, Mass Spectrometry, Molecular Sequence Data, Nervous System, Neurons, Nonmammalian Embryo, Proteasome Endopeptidase Complex, Proteomics, Reproducibility of Results, Ubiquitin, Ubiquitination, Western Blotting, Animals, Drosophila melanogaster |
| Source: | PLoS ONE |
| ISSN: | 1932-6203 |
| Publisher: | Public Library of Science |
| Volume: | 10 |
| Number: | 10 |
| Page Range: | e0139083 |
| Date: | 13 October 2015 |
| Official Publication: | https://doi.org/10.1371/journal.pone.0139083 |
| PubMed: | View item in PubMed |
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