A genome-wide genetic screen identifies a novel kDNA replication protein in trypanosomes.
Publisher DOI
PubMed ID
42165136
Abstract
Mitochondrial DNA of trypanosomatid parasites is organized into a topologically complex structure, named kinetoplast [kinetoplast DNA (kDNA)]. Replication, segregation and expression of kDNA involve an estimated ∼300 proteins, only a fraction of which have been identified and characterized. Here, we report the development of a genetic screen in Trypanosoma brucei to identify novel kDNA maintenance factors. Of the 20 highest-ranked genes identified, six are known kDNA maintenance factors. We selected one hit, Tb927.8.4240, a gene of previously unknown function, for further experimental characterization. Ultrastructure expansion microscopy using a tagged version of the protein reveals a dynamic localization during the cell cycle. RNA interference-mediated ablation of Tb927.8.4240 results in the progressive but incomplete loss of kDNA, with only a minor effect on the tripartite attachment complex, suggesting the protein is involved in kDNA replication but not segregation. The growth phenotype of Tb927.8.4240 ablation is fully rescued in a kDNA-independent genetic background, confirming a specific role in kDNA replication. In summary, we describe a functional genetic screen for the identification of kDNA maintenance factors in trypanosomes, validate one hit as a novel kDNA replication factor, and provide a prioritized hit list as a promising starting point for the future identification of additional factors.
Date Issued
2026-05-20
Publication Type
Article
Subject(s)
Language(s)
en
Author(s)
Miskinyte, Migla | |
Ivens, Alasdair | |
Waterfall, Martin | |
Gould, Matthew K | |
Glover, Lucy | |
Horn, David | |
Schnaufer, Achim |
Journal
Nucleic Acids Research
Publisher
Oxford University Press
ISSN
1362-4962
0305-1048
Access(Rights)
open.access