Essential roles for deubiquitination in Leishmania life cycle progression

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:Rodenko, Dr Boris and Damianou, Andreas and Burchmore, Dr Richard and Geoghegan, Vincent 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 > Institute of Infection Immunity and Inflammation
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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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
168381Proteolysis and life cycle progression in LeishmaniaJeremy MottramMedical Research Council (MRC)MR/K019384/1III - Parasitology