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Inorganic Polyphosphate Modulates Chromosome Transmission Fidelity in the Fission Yeast <i>Schizosaccharomyces pombe</i>.

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TUG protein acts through a disordered region to organize the early secretory pathway.

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Inositol Pyrophosphate-Controlled Kinetochore Architecture and Mitotic Entry in <i>S. pombe</i>.

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The PPIP5K Family Member Asp1 Controls Inorganic Polyphosphate Metabolism in <i>S. pombe</i>.

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Related Experiment Video

Updated: May 8, 2026

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay PCA in Living Cells
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An In Vivo Yeast-Based Activity Assay for the PPIP5K Family.

Abel R Alcázar-Román1, Ursula Fleig2

  • 1Eukaryotic Microbiology, Heinrich-Heine-University, Düsseldorf, Germany. abel.alcazar@hhu.de.

Methods in Molecular Biology (Clifton, N.J.)
|August 29, 2025
PubMed
Summary

We developed a fast in vivo assay in fission yeast to assess the kinase and pyrophosphatase activities of PPIP5K enzymes. This method aids in understanding cellular inositol polyphosphate levels and their biological roles.

Keywords:
1,5-InsP8Asp1In VivoMicrotubulesPPIP5KSchizosaccharomyces pombeTBZThiabendazoleYeast

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • The PPIP5K enzyme family is crucial for regulating cellular inositol polyphosphate levels, specifically 1,5-InsP8.
  • PPIP5K enzymes possess dual kinase and pyrophosphatase activities, essential for maintaining cellular homeostasis.
  • Understanding these bifunctional activities is vital for elucidating diverse biological processes regulated by 1,5-InsP8.

Purpose of the Study:

  • To develop a novel, rapid, and efficient in vivo assay for assessing PPIP5K enzyme activity.
  • To provide a complementary method to biochemical assays for functional characterization of PPIP5K family members across species.
  • To facilitate research into the biological roles of 1,5-InsP8 by enabling easier study of its regulating enzymes.

Main Methods:

  • Utilized the fission yeast Schizosaccharomyces pombe as a model organism for the in vivo assay.
  • Developed a functional assay to simultaneously assess both the kinase and pyrophosphatase activities of PPIP5K enzymes.
  • The assay is designed for simplicity and speed, allowing for high-throughput screening or rapid functional assessment.

Main Results:

  • Successfully established a simple and fast in vivo assay for evaluating PPIP5K bifunctional activities.
  • Demonstrated the utility of the assay for functional assessment of PPIP5K family members from various species.
  • The assay provides a complementary approach to traditional biochemical methods for studying enzyme function.

Conclusions:

  • The developed fission yeast assay offers a valuable tool for the functional characterization of PPIP5K enzymes.
  • This assay will accelerate research into the roles of 1,5-InsP8 in cellular processes.
  • The method's simplicity and speed make it broadly applicable for studying PPIP5K orthologs in diverse eukaryotic organisms.