Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single cell transcriptomics

Briggs, E. M., Marques, C. A. , Oldrieve, G. R., Hu, J., Otto, T. D. and Matthews, K. R. (2023) Profiling the bloodstream form and procyclic form Trypanosoma brucei cell cycle using single cell transcriptomics. eLife, 12, e86325. (doi: 10.7554/eLife.86325) (PMID:37166108) (PMCID:PMC10212563)

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Abstract

African trypanosomes proliferate as bloodstream forms and procyclic forms in the mammal and tsetse fly midgut, respectively. This allows them to colonise the host environment upon infection and ensure life cycle progression. Yet, understanding of the mechanisms that regulate and drive the cell replication cycle of these forms is limited. Using single cell transcriptomics on unsynchronised cell populations, we have obtained high resolution cell cycle regulated transcriptomes of both procyclic and slender bloodstream form Trypanosoma brucei without prior cell sorting or synchronisation. Additionally, we describe an efficient freeze-thawing protocol that allows single cell transcriptomic analysis of cryopreserved T. brucei. Computational reconstruction of the cell cycle using periodic pseudotime inference allowed the dynamic expression patterns of cycling genes to be profiled for both life cycle forms. Comparative analyses identify a core cycling transcriptome highly conserved between forms, as well as several genes where transcript levels dynamics are form-specific. Comparing transcript expression patterns with protein abundance revealed that the majority of genes with periodic cycling transcript and protein levels exhibit a relative delay between peak transcript and protein expression. This work reveals novel detail of the cell cycle regulated transcriptomes of both forms, which are available for further interrogation via an interactive webtool.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Briggs, Miss Emma and Otto, Professor Thomas and De Almeida Marques, Dr Catarina
Authors: Briggs, E. M., Marques, C. A., Oldrieve, G. R., Hu, J., Otto, T. D., and Matthews, K. R.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity
Research Centre:College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Immunobiology
Journal Name:eLife
Publisher:eLife Sciences Publications
ISSN:2050-084X
ISSN (Online):2050-084X
Published Online:11 May 2023
Copyright Holders:Copyright © 2023 Briggs et al.
First Published:First published in eLife 12: e86325
Publisher Policy:Reproduced under a Creative Commons License
Data DOI:10.5281/zenodo.7508131

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
170547The Wellcome Centre for Molecular Parasitology ( Core Support )Andrew WatersWellcome Trust (WELLCOTR)104111/Z/14/ZIII - Parasitology
301963Does genome replication in the protozoan parasite Leishmania rely on origin-indepenent initiation?Richard McCullochBiotechnology and Biological Sciences Research Council (BBSRC)BB/R017166/1III - Parasitology
313882A distinct mode of DNA replication initiation in trypanosomes?Richard McCullochBiotechnology and Biological Sciences Research Council (BBSRC)BB/W001101/1III - Parasitology