Damianou, A., Burge, R. J., Catta-Preta, C. M.C., Geoghegan, V., Nievas, Y. R., Newling, K., Brown, E., Burchmore, R. , Rodenko, B. and Mottram, J. C. (2020) Essential roles for deubiquitination in Leishmania life cycle progression. PLoS Pathogens, 16(6), e1008455. (doi: 10.1371/journal.ppat.1008455) (PMID:32544189) (PMCID:PMC7319358)
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Abstract
The parasitic protozoan Leishmania requires proteasomal, autophagic and lysosomal proteolytic pathways to enact the extensive cellular remodelling that occurs during its life cycle. The proteasome is essential for parasite proliferation, yet little is known about the requirement for ubiquitination/deubiquitination processes in growth and differentiation. Activity-based protein profiling of L. mexicana C12, C19 and C65 deubiquitinating cysteine peptidases (DUBs) revealed DUB activity remains relatively constant during differentiation of procyclic promastigote to amastigote. However, when life cycle phenotyping (bar-seq) was performed on a pool including 15 barcoded DUB null mutants created in promastigotes using CRISPR-Cas9, significant loss of fitness was observed during differentiation and intracellular infection. DUBs 4, 7, and 13 are required for successful transformation from metacyclic promastigote to amastigote and DUBs 3, 5, 6, 8, 10, 11 and 14 are required for normal amastigote proliferation in mice. DUBs 1, 2, 12 and 16 are essential for promastigote viability and the essential role of DUB2 in establishing infection was demonstrated using DiCre inducible gene deletion in vitro and in vivo. DUB2 is found in the nucleus and interacts with nuclear proteins associated with transcription/chromatin dynamics, mRNA splicing and mRNA capping. DUB2 has broad linkage specificity, cleaving all the di-ubiquitin chains except for Lys27 and Met1. Our study demonstrates the crucial role that DUBs play in differentiation and intracellular survival of Leishmania and that amastigotes are exquisitely sensitive to disruption of ubiquitination homeostasis.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Burchmore, Dr Richard and Geoghegan, Vincent and Rodenko, Dr Boris and Damianou, Andreas and Mottram, Professor Jeremy |
| Creator Roles: | Damianou, A.Conceptualization, Investigation, Visualization, Writing – original draft, Writing – review and editing Geoghegan, V.Conceptualization, Data curation, Investigation, Resources, Writing – original draft, Writing – review and editing Burchmore, R.Supervision, Writing – review and editing Rodenko, B.Conceptualization, Funding acquisition, Supervision, Writing – review and editing Mottram, J. C.Conceptualization, Funding acquisition, Supervision, Writing – original draft, Writing – review and editing |
| Authors: | Damianou, A., Burge, R. J., Catta-Preta, C. M.C., Geoghegan, V., Nievas, Y. R., Newling, K., Brown, E., Burchmore, R., Rodenko, B., and Mottram, J. C. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Infection & Immunity |
| Journal Name: | PLoS Pathogens |
| Publisher: | Public Library of Science |
| ISSN: | 1553-7366 |
| ISSN (Online): | 1553-7374 |
| Published Online: | 16 June 2020 |
| Copyright Holders: | Copyright © 2020 Damianou et al. |
| First Published: | First published in PLoS Pathogens 16(6): e1008455 |
| Publisher Policy: | Reproduced under a Creative Commons License |
| Data DOI: | 10.25345/C5Z10J |
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