Conditional genome engineering reveals canonical and divergent roles for the Hus1 component of the 9-1-1 complex in the maintenance of the plastic genome of Leishmania

Damasceno, J. D., Obonaga, R., Silva, G. L.A., Reis-Cunha, J. L., Duncan, S. M., Bartholomeu, D. C., Mottram, J. C. , McCulloch, R. and Tosi, L. R.O. (2018) Conditional genome engineering reveals canonical and divergent roles for the Hus1 component of the 9-1-1 complex in the maintenance of the plastic genome of Leishmania. Nucleic Acids Research, 46(22), pp. 11835-11846. (doi: 10.1093/nar/gky1017) (PMID:30380080) (PMCID:PMC6294564)

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Abstract

Leishmania species are protozoan parasites whose remarkably plastic genome limits the establishment of effective genetic manipulation and leishmaniasis treatment. The strategies used by Leishmania to maintain its genome while allowing variability are not fully understood. Here, we used DiCre-mediated conditional gene deletion to show that HUS1, a component of the 9–1–1 (RAD9-RAD1-HUS1) complex, is essential and is required for a G2/M checkpoint. By analyzing genome-wide instability in HUS1 ablated cells, HUS1 is shown to have a conserved role, by which it preserves genome stability and also a divergent role, by which it promotes genome variability. These roles of HUS1 are related to distinct patterns of formation and resolution of single-stranded DNA and γH2A, throughout the cell cycle. Our findings suggest that Leishmania 9–1–1 subunits have evolved to co-opt canonical genomic maintenance and genomic variation functions. Hence, this study reveals a pivotal function of HUS1 in balancing genome stability and transmission in Leishmania. These findings may be relevant to understanding the evolution of genome maintenance and plasticity in other pathogens and eukaryotes.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Duncan, Mr Samuel and Damasceno, Dr Jeziel and McCulloch, Professor Richard and Mottram, Professor Jeremy
Authors: Damasceno, J. D., Obonaga, R., Silva, G. L.A., Reis-Cunha, J. L., Duncan, S. M., Bartholomeu, D. C., Mottram, J. C., McCulloch, R., and Tosi, L. R.O.
College/School:College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Nucleic Acids Research
Publisher:Oxford University Press
ISSN:0305-1048
ISSN (Online):1362-4962
Published Online:31 October 2018
Copyright Holders:Copyright © 2018 The Authors
First Published:First published in Nucleic Acids Research 46(22): 11835-11846
Publisher Policy:Reproduced under a Creative Commons License

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