The TOR pathway modulates cytoophidium formation in Schizosaccharomyces pombe

J Biol Chem. 2019 Oct 4;294(40):14686-14703. doi: 10.1074/jbc.RA119.009913. Epub 2019 Aug 19.

Abstract

CTP synthase (CTPS) has been demonstrated to form evolutionarily-conserved filamentous structures termed cytoophidia whose exact cellular functions remain unclear, but they may play a role in intracellular compartmentalization. We have previously shown that the mammalian target of rapamycin complex 1 (mTORC1)-S6K1 pathway mediates cytoophidium assembly in mammalian cells. Here, using the fission yeast Schizosaccharomyces pombe as a model of a unicellular eukaryote, we demonstrate that the target of rapamycin (TOR)-signaling pathway regulates cytoophidium formation (from the S. pombe CTPS ortholog Cts1) also in S. pombe Conducting a systematic analysis of all viable single TOR subunit-knockout mutants and of several major downstream effector proteins, we found that Cts1 cytoophidia are significantly shortened and often dissociate when TOR is defective. We also found that the activities of the downstream effector kinases of the TORC1 pathway, Sck1, Sck2, and Psk1 S6, as well as of the S6K/AGC kinase Gad8, the major downstream effector kinase of the TORC2 pathway, are necessary for proper cytoophidium filament formation. Interestingly, we observed that the Crf1 transcriptional corepressor for ribosomal genes is a strong effector of Cts1 filamentation. Our findings connect TOR signaling, a major pathway required for cell growth, with the compartmentalization of the essential nucleotide synthesis enzyme CTPS, and we uncover differences in the regulation of its filamentation among higher multicellular and unicellular eukaryotic systems.

Keywords: CTP synthase; Schizosaccharomyces pombe; TOR complex (TORC); cytoophidium; fission yeast; kinase signaling; metabolic regulation; nucleotide; protein filamentation; pyrimidine; signal transduction; subcellular organelle; target of rapamycin (TOR); yeast; yeast genetics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carbon-Nitrogen Ligases / chemistry
  • Carbon-Nitrogen Ligases / genetics*
  • Cell Compartmentation / genetics
  • Cytoplasm / genetics
  • Gene Knockout Techniques
  • Mechanistic Target of Rapamycin Complex 1 / genetics*
  • Mechanistic Target of Rapamycin Complex 2 / genetics
  • Phosphorylation / genetics
  • Protein Serine-Threonine Kinases / chemistry
  • Protein Serine-Threonine Kinases / genetics
  • Receptors, Corticotropin-Releasing Hormone / chemistry
  • Receptors, Corticotropin-Releasing Hormone / genetics*
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces pombe Proteins / chemistry
  • Schizosaccharomyces pombe Proteins / genetics
  • Signal Transduction / genetics
  • Transcription Factors / chemistry
  • Transcription Factors / genetics

Substances

  • Receptors, Corticotropin-Releasing Hormone
  • Schizosaccharomyces pombe Proteins
  • Transcription Factors
  • CRF receptor type 1
  • Sck2 protein, S pombe
  • Mechanistic Target of Rapamycin Complex 1
  • Mechanistic Target of Rapamycin Complex 2
  • Protein Serine-Threonine Kinases
  • Carbon-Nitrogen Ligases
  • CTP synthetase