Wallbank, BA; Dominicus, CS; Broncel, M; Legrave, N; MacRae, JI; Staines, HM; Treeck, M
(2019)
Characterisation of the Toxoplasma gondii tyrosine transporter and its phosphorylation by the calcium-dependent protein kinase 3.
Mol Microbiol, 111 (5).
pp. 1167-1181.
ISSN 1365-2958
https://doi.org/10.1111/mmi.14156
SGUL Authors: Staines, Henry Michael
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Abstract
Toxoplasma gondii parasites rapidly exit their host cell when exposed to calcium ionophores. Calcium-dependent protein kinase 3 (TgCDPK3) was previously identified as a key mediator in this process, as TgCDPK3 knockout (∆cdpk3) parasites fail to egress in a timely manner. Phosphoproteomic analysis comparing WT with ∆cdpk3 parasites revealed changes in the TgCDPK3-dependent phosphoproteome that included proteins important for regulating motility, but also metabolic enzymes, indicating that TgCDPK3 controls processes beyond egress. Here we have investigated a predicted direct target of TgCDPK3, ApiAT5-3, a putative transporter of the major facilitator superfamily, and show that it is rapidly phosphorylated at serine 56 after induction of calcium signalling. Conditional knockout of apiAT5-3 results in transcriptional upregulation of most ribosomal subunits, but no alternative transporters, and subsequent parasite death. Mutating the S56 to a non-phosphorylatable alanine leads to a fitness cost, suggesting that phosphorylation of this residue is beneficial, albeit not essential, for tyrosine import. Using a combination of metabolomics and heterologous expression, we confirmed a primary role in tyrosine import for ApiAT5-3. However, no significant differences in tyrosine import could be detected in phosphorylation site mutants showing that if tyrosine transport is affected by S56 phosphorylation, its regulatory role is subtle.
Item Type: | Article | |||||||||||||||||||||
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Additional Information: | © 2018 The Authors. Molecular Microbiology Published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits use, distribution and reproduction in any medium, provided the original work is properly cited. | |||||||||||||||||||||
Keywords: | 06 Biological Sciences, 11 Medical And Health Sciences, 07 Agricultural And Veterinary Sciences, Microbiology | |||||||||||||||||||||
SGUL Research Institute / Research Centre: | Academic Structure > Infection and Immunity Research Institute (INII) | |||||||||||||||||||||
Journal or Publication Title: | Mol Microbiol | |||||||||||||||||||||
ISSN: | 1365-2958 | |||||||||||||||||||||
Language: | eng | |||||||||||||||||||||
Publisher License: | Creative Commons: Attribution 4.0 | |||||||||||||||||||||
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PubMed ID: | 30402958 | |||||||||||||||||||||
Go to PubMed abstract | ||||||||||||||||||||||
URI: | https://openaccess.sgul.ac.uk/id/eprint/110308 | |||||||||||||||||||||
Publisher's version: | https://doi.org/10.1111/mmi.14156 |
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