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The Protein Neddylation Pathway in Trypanosoma brucei
- Source :
- Journal of Biological Chemistry. 292:1081-1091
- Publication Year :
- 2017
- Publisher :
- Elsevier BV, 2017.
-
Abstract
- Protein posttranslational modifications such as neddylation play crucial roles in regulating protein function. Only a few neddylated substrates have been validated to date, and the role of neddylation remains poorly understood. Here, using Trypanosoma brucei as the model organism, we investigated the function and substrates of TbNedd8. TbNedd8 is distributed throughout the cytosol but enriched in the nucleus and the flagellum. Depletion of TbNedd8 by RNAi abolished global protein ubiquitination, caused DNA re-replication in postmitotic cells, impaired spindle assembly, and compromised the flagellum attachment zone filament, leading to flagellum detachment. Through affinity purification and mass spectrometry, we identified 70 TbNedd8-conjugated and -associated proteins, including known Nedd8-conjugated and -associated proteins, putative TbNedd8 conjugation system enzymes, proteins of diverse biological functions, and proteins of unknown function. Finally, we validated six Cullins as bona fide TbNedd8 substrates and identified the TbNedd8 conjugation site in three Cullins. This work lays the foundation for understanding the roles of protein neddylation in this early divergent parasitic protozoan.
- Subjects :
- 0301 basic medicine
030102 biochemistry & molecular biology
biology
Cullin Proteins
Cell Biology
Trypanosoma brucei
Flagellum
biology.organism_classification
Biochemistry
NEDD8
Protein ubiquitination
Cell biology
03 medical and health sciences
Protein neddylation
030104 developmental biology
biology.protein
Neddylation
Molecular Biology
Cullin
Subjects
Details
- ISSN :
- 00219258
- Volume :
- 292
- Database :
- OpenAIRE
- Journal :
- Journal of Biological Chemistry
- Accession number :
- edsair.doi...........665d932d681619873d2cbc88b861a20d