Abstract
The Toxoplasma genome encodes the capacity for distinct architectures underlying cell cycle progression in a life cycle stage-dependent manner. Replication in intermediate hosts occurs by endodyogeny, whereas a hybrid of schizogony and endopolygeny occurs in the gut of the definitive feline host. Here, we characterize the consequence of the loss of a cell cycle-regulated ovarian tumor (OTU family) deubiquitinase, OTUD3A of Toxoplasma gondii (TgOTUD3A; TGGT1_258780), in T. gondii tachyzoites. Rather than the mutation being detrimental, mutant parasites exhibited a fitness advantage, outcompeting the wild type. This phenotype was due to roughly one-third of TgOTUD3A-knockout (TgOTUD3A-KO) tachyzoites exhibiting deviations from endodyogeny by employing replication strategies that produced 3, 4, or 5 viable progeny within a gravid mother instead of the usual 2. We established the mechanistic basis underlying these altered replication strategies to be a dysregulation of centrosome duplication, causing a transient loss of stoichiometry between the inner and outer cores that resulted in a failure to terminate S phase at the attainment of 2N ploidy and/or the decoupling of mitosis and cytokinesis. The resulting dysregulation manifested as deviations in the normal transitions from S phase to mitosis (S/M) (endopolygeny-like) or M phase to cytokinesis (M/C) (schizogony-like). Notably, these imbalances are corrected prior to cytokinesis, resulting in the generation of normal progeny. Our findings suggest that decisions regarding the utilization of specific cell cycle architectures are controlled by a ubiquitin-mediated mechanism that is dependent on the absolute threshold levels of an as-yet-unknown target(s). Analysis of the TgOTUD3A-KO mutant provides new insights into mechanisms underlying the plasticity of apicomplexan cell cycle architecture.
Document Type
Article
Publication Date
11-21-2017
Digital Object Identifier (DOI)
https://doi.org/10.1128/mBio.01846-17
Funding Information
This work was supported by Kentucky Science and Engineering Foundation grant number KSEF-2624-RDE-015 and National Institutes of Health grant number NIH R21 AI122894 (both awarded to A.P.S.).
Related Content
Supplemental material for this article may be found at https://doi.org/10.1128/mBio.01846-17.
Repository Citation
Dhara, Animesh; de Paula Baptista, Rodrigo; Kissinger, Jessica C.; Snow, Ernest Charles; and Sinai, Anthony P., "Ablation of an Ovarian Tumor Family Deubiquitinase Exposes the Underlying Regulation Governing the Plasticity of Cell Cycle Progression in Toxoplasma gondii" (2017). Microbiology, Immunology, and Molecular Genetics Faculty Publications. 96.
https://uknowledge.uky.edu/microbio_facpub/96
TEXT S1
inline-supplementary-material-1.tif (184 kB)
FIG S1
inline-supplementary-material-2.tif (421 kB)
FIG S2
inline-supplementary-material-3.tif (339 kB)
FIG S3
inline-supplementary-material-4.tif (458 kB)
FIG S4
inline-supplementary-material-5.tif (282 kB)
FIG S5
inline-supplementary-material-6.tif (631 kB)
FIG S6
inline-supplementary-material-8.tif (13191 kB)
FIG S7
inline-supplementary-material-9.pdf (113 kB)
TABLE S1
Notes/Citation Information
Published in mBio, v. 8, issue 6, e01846-17, p. 1-28.
Copyright © 2017 Dhara et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.