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CTPS forms the cytoophidium in zebrafish.

Chia-Chun Chang1, Gerson Dierley Keppeke2, Christopher L Antos1

  • 1School of Life Science and Technology, ShanghaiTech University, Shanghai, 201210, China.

Experimental Cell Research
|June 15, 2021
PubMed
Summary

Zebrafish cytidine triphosphate synthase (CTPS) forms intracellular structures called cytoophidia. These structures are naturally present and can be modulated by inhibitors, suggesting physiological roles.

Keywords:
CTPSCytoophidiumZebrafish

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Developmental Biology

Background:

  • Cytidine triphosphate synthase (CTPS) is crucial for de novo CTP biosynthesis, catalyzing a rate-limiting step.
  • Intracellular CTPS structures, known as cytoophidia, are observed in various organisms and may regulate CTPS activity and stability.
  • The physiological significance of cytoophidia in vertebrates remains largely unexplored.

Purpose of the Study:

  • To investigate the ability of zebrafish CTPS1a and CTPS1b to form cytoophidia in cultured cells.
  • To determine the presence and distribution of CTPS cytoophidia in zebrafish tissues under normal and induced conditions.
  • To elucidate the role of specific mutations and inhibitors in cytoophidium assembly.

Main Methods:

  • Expression of zebrafish CTPS1a and CTPS1b in cultured cells.
  • Site-directed mutagenesis to create the H355A mutant.
  • Histological analysis of CTPS cytoophidia in various zebrafish tissues.
  • Treatment with the CTPS inhibitor 6-diazo-5-oxo-l-norleucine (DON).

Main Results:

  • Both zebrafish CTPS1a and CTPS1b can form cytoophidia in cultured cells.
  • A specific point mutation (H355A) in CTPS disrupts cytoophidium assembly.
  • CTPS cytoophidia are naturally present in multiple tissues of larval and adult zebrafish.
  • Treatment with DON increases cytoophidium formation in certain zebrafish tissues.

Conclusions:

  • Cytoophidium formation is a natural phenomenon in zebrafish CTPS.
  • The H355A mutation highlights the importance of specific residues for cytoophidium assembly.
  • Modulation of cytoophidia by inhibitors suggests their involvement in physiological processes.
  • These findings provide a foundation for further research into the function of CTPS cytoophidia in vertebrates.