TgPRELID, a mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii

Victoria Jeffers, Edwin T. Kamau, Ananth R. Srinivasan, Jonathan Harper, Preethi Sankaran, Sarah E. Post, Joseph M. Varberg, William Sullivan, Jon P. Boyle

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

New drugs to control infection with the protozoan parasite Toxoplasma gondii are needed as current treatments exert toxic side effects on patients. Approaches to develop novel compounds for drug development include screening of compound libraries and targeted inhibition of essential cellular pathways. We identified two distinct compounds that display inhibitory activity against the parasite's replicative stage: F3215-0002, which we previously identified during a compound library screen, and I-BET151, an inhibitor of bromodomains, the "reader" module of acetylated lysines. In independent studies, we sought to determine the targets of these two compounds using forward genetics, generating resistant mutants and identifying the determinants of resistance with comparative genome sequencing. Despite the dissimilarity of the two compounds, we recovered resistant mutants with nonsynonymous mutations in the same domain of the same gene, TGGT1_254250, which we found encodes a protein that localizes to the parasite mitochondrion (designated TgPRELID after the name of said domain). We found that mutants selected with one compound were cross resistant to the other compound, suggesting a common mechanism of resistance. To further support our hypothesis that TgPRELID mutations facilitate resistance to both I-BET151 and F3215-0002, CRISPR (clustered regularly interspaced short palindromic repeat)/CAS9-mediated mutation of TgPRELID directly led to increased F3215-0002 resistance. Finally, all resistance mutations clustered in the same subdomain of TgPRELID. These findings suggest that TgPRELID may encode a multidrug resistance factor or that I-BET151 and F3215-0002 have the same target(s) despite their distinct chemical structures.

Original languageEnglish (US)
Article numbere00229-16
JournalmSphere
Volume2
Issue number1
DOIs
StatePublished - Jan 1 2017

Fingerprint

Mitochondrial Proteins
Toxoplasma
Multiple Drug Resistance
Parasites
Mutation
Libraries
Clustered Regularly Interspaced Short Palindromic Repeats
Protozoan Infections
R Factors
Poisons
Drug and Narcotic Control
Lysine
Names
Mitochondria
Genome
Pharmaceutical Preparations
Genes
GSK1210151A
Proteins
Therapeutics

Keywords

  • Mitochondrial protein import
  • Multidrug resistance
  • PRELI domain
  • Toxoplasma gondii

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Jeffers, V., Kamau, E. T., Srinivasan, A. R., Harper, J., Sankaran, P., Post, S. E., ... Boyle, J. P. (2017). TgPRELID, a mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii. mSphere, 2(1), [e00229-16]. https://doi.org/10.1128/mSphere.00229-16

TgPRELID, a mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii. / Jeffers, Victoria; Kamau, Edwin T.; Srinivasan, Ananth R.; Harper, Jonathan; Sankaran, Preethi; Post, Sarah E.; Varberg, Joseph M.; Sullivan, William; Boyle, Jon P.

In: mSphere, Vol. 2, No. 1, e00229-16, 01.01.2017.

Research output: Contribution to journalArticle

Jeffers, V, Kamau, ET, Srinivasan, AR, Harper, J, Sankaran, P, Post, SE, Varberg, JM, Sullivan, W & Boyle, JP 2017, 'TgPRELID, a mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii', mSphere, vol. 2, no. 1, e00229-16. https://doi.org/10.1128/mSphere.00229-16
Jeffers V, Kamau ET, Srinivasan AR, Harper J, Sankaran P, Post SE et al. TgPRELID, a mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii. mSphere. 2017 Jan 1;2(1). e00229-16. https://doi.org/10.1128/mSphere.00229-16
Jeffers, Victoria ; Kamau, Edwin T. ; Srinivasan, Ananth R. ; Harper, Jonathan ; Sankaran, Preethi ; Post, Sarah E. ; Varberg, Joseph M. ; Sullivan, William ; Boyle, Jon P. / TgPRELID, a mitochondrial protein linked to multidrug resistance in the parasite Toxoplasma gondii. In: mSphere. 2017 ; Vol. 2, No. 1.
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abstract = "New drugs to control infection with the protozoan parasite Toxoplasma gondii are needed as current treatments exert toxic side effects on patients. Approaches to develop novel compounds for drug development include screening of compound libraries and targeted inhibition of essential cellular pathways. We identified two distinct compounds that display inhibitory activity against the parasite's replicative stage: F3215-0002, which we previously identified during a compound library screen, and I-BET151, an inhibitor of bromodomains, the {"}reader{"} module of acetylated lysines. In independent studies, we sought to determine the targets of these two compounds using forward genetics, generating resistant mutants and identifying the determinants of resistance with comparative genome sequencing. Despite the dissimilarity of the two compounds, we recovered resistant mutants with nonsynonymous mutations in the same domain of the same gene, TGGT1_254250, which we found encodes a protein that localizes to the parasite mitochondrion (designated TgPRELID after the name of said domain). We found that mutants selected with one compound were cross resistant to the other compound, suggesting a common mechanism of resistance. To further support our hypothesis that TgPRELID mutations facilitate resistance to both I-BET151 and F3215-0002, CRISPR (clustered regularly interspaced short palindromic repeat)/CAS9-mediated mutation of TgPRELID directly led to increased F3215-0002 resistance. Finally, all resistance mutations clustered in the same subdomain of TgPRELID. These findings suggest that TgPRELID may encode a multidrug resistance factor or that I-BET151 and F3215-0002 have the same target(s) despite their distinct chemical structures.",
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