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Identification of

Agnieszka Lis1, Carlos Gustavo Baptista1, Kelsey Dahlgren1

  • 1Department of Microbiology and Immunology, SUNY at Buffalo School of Medicine, Buffalo, New York, USA.

Msphere
|June 5, 2023
PubMed
Summary
This summary is machine-generated.

Toxoplasma gondii uses CDPK3 as an extracellular stress-induced eEF2 kinase to enhance protein synthesis during extracellular survival. This kinase relocates to the cytosol to interact with eEF2, preventing its phosphorylation.

Keywords:
Toxoplasma gondiikinaseoxygenparasiteprotein synthesis

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Area of Science:

  • Parasitology
  • Molecular Biology
  • Cellular Biology

Background:

  • Toxoplasma gondii tachyzoites require stress response pathways for extracellular survival.
  • Protein synthesis regulation is crucial for parasite adaptation to extracellular environments.
  • Previous work identified PHYb's role in regulating protein elongation factor eEF2.

Purpose of the Study:

  • To identify the eEF2 kinase in Toxoplasma.
  • To investigate the role of CDPK3 in the parasite's extracellular stress response.
  • To elucidate the mechanism of CDPK3-eEF2 interaction under stress.

Main Methods:

  • BLAST analysis to identify homologous kinases.
  • Gene deletion studies to assess CDPK3 function.
  • Co-immunoprecipitation to study protein interactions.
  • Subcellular localization studies (N-myristoylation).

Main Results:

  • CDPK3 was identified as the most homologous kinase to yeast eEF2 kinase.
  • Loss of CDPK3 resulted in decreased eEF2 phosphorylation and increased elongation rates.
  • CDPK3 and eEF2 interact in stressed parasites.
  • Under stress, CDPK3 relocalizes to the cytosol, suggesting a mechanism for interaction.

Conclusions:

  • CDPK3 is the first identified protozoan extracellular stress-induced eEF2 kinase.
  • CDPK3 plays a critical role in regulating protein synthesis during Toxoplasma extracellular stress.
  • CDPK3's cytoplasmic relocalization is key to its interaction with eEF2.