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Genome-scale functional profiling of cell cycle controls in African trypanosomes

View ORCID ProfileCatarina A. Marques, View ORCID ProfileMichele Tinti, Andrew Cassidy, View ORCID ProfileDavid Horn
doi: https://doi.org/10.1101/2020.07.16.206698
Catarina A. Marques
1Wellcome Trust Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, United Kingdom
3Wellcome Trust Centre for Integrative Parasitology, University of Glasgow, United Kingdom
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Michele Tinti
1Wellcome Trust Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, United Kingdom
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Andrew Cassidy
2Tayside Centre for Genomic Analysis, Ninewells Hospital and School of Medicine, University of Dundee, United Kingdom
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David Horn
1Wellcome Trust Centre for Anti-Infectives Research, School of Life Sciences, University of Dundee, United Kingdom
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  • For correspondence: d.horn@dundee.ac.uk
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Abstract

Trypanosomatids, which include major pathogens of humans and livestock, are divergent eukaryotes for which cell cycle controls and the underlying mechanisms are not completely understood. Here, we describe a genome-wide RNA-interference library screen for cell cycle regulators in bloodstream form Trypanosoma brucei. We induced massive parallel knockdown and sorted the perturbed population into cell cycle stages using flow cytometry. RNAi-targets were deep-sequenced from each stage and cell cycle profiles were digitally reconstructed at a genomic scale. We identify hundreds of proteins that impact cell cycle progression; glycolytic enzymes required for G1S progression, DNA replication factors, mitosis regulators, proteasome and kinetochore complex components required for G2M progression, flagellar and cytoskeletal components required for cytokinesis, mRNA-binding factors, protein kinases and many previously uncharacterised proteins. The outputs facilitate functional annotation and drug-target prioritisation and provide comprehensive functional genomic evidence for the machineries, pathways and regulators that coordinate progression through the trypanosome cell cycle.

The data can be searched and browsed using an interactive, open access, online data visualization tool (https://tryp-cycle.onrender.com).

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • ↵§ joint first authors

  • https://tryp-cycle.onrender.com

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted July 16, 2020.
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Genome-scale functional profiling of cell cycle controls in African trypanosomes
Catarina A. Marques, Michele Tinti, Andrew Cassidy, David Horn
bioRxiv 2020.07.16.206698; doi: https://doi.org/10.1101/2020.07.16.206698
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Genome-scale functional profiling of cell cycle controls in African trypanosomes
Catarina A. Marques, Michele Tinti, Andrew Cassidy, David Horn
bioRxiv 2020.07.16.206698; doi: https://doi.org/10.1101/2020.07.16.206698

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