Specific detection of fission yeast primary septum reveals septum and cleavage furrow ingression during early anaphase independent of mitosis completion

PLoS Genet. 2018 May 29;14(5):e1007388. doi: 10.1371/journal.pgen.1007388. eCollection 2018 May.

Abstract

It is widely accepted in eukaryotes that the cleavage furrow only initiates after mitosis completion. In fission yeast, cytokinesis requires the synthesis of a septum tightly coupled to cleavage furrow ingression. The current cytokinesis model establishes that simultaneous septation and furrow ingression only initiate after spindle breakage and mitosis exit. Thus, this model considers that although Cdk1 is inactivated at early-anaphase, septation onset requires the long elapsed time until mitosis completion and full activation of the Hippo-like SIN pathway. Here, we studied the precise timing of septation onset regarding mitosis by exploiting both the septum-specific detection with the fluorochrome calcofluor and the high-resolution electron microscopy during anaphase and telophase. Contrarily to the existing model, we found that both septum and cleavage furrow start to ingress at early anaphase B, long before spindle breakage, with a slow ingression rate during anaphase B, and greatly increasing after telophase onset. This shows that mitosis and cleavage furrow ingression are not concatenated but simultaneous events in fission yeast. We found that the timing of septation during early anaphase correlates with the cell size and is regulated by the corresponding levels of SIN Etd1 and Rho1. Cdk1 inactivation was directly required for timely septation in early anaphase. Strikingly the reduced SIN activity present after Cdk1 loss was enough to trigger septation by immediately inducing the medial recruitment of the SIN kinase complex Sid2-Mob1. On the other hand, septation onset did not depend on the SIN asymmetry establishment, which is considered a hallmark for SIN activation. These results recalibrate the timing of key cytokinetic events in fission yeast; and unveil a size-dependent control mechanism that synchronizes simultaneous nuclei separation with septum and cleavage furrow ingression to safeguard the proper chromosome segregation during cell division.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anaphase / physiology*
  • Benzenesulfonates / chemistry
  • CDC2 Protein Kinase / physiology
  • Cell Cycle Proteins / physiology*
  • Cell Nucleus / physiology
  • Cytokinesis / physiology*
  • Microscopy, Electron, Transmission
  • Microscopy, Fluorescence / methods
  • Protein Kinases / physiology
  • Schizosaccharomyces / physiology*
  • Schizosaccharomyces / ultrastructure
  • Schizosaccharomyces pombe Proteins / physiology*
  • Spindle Apparatus / physiology*
  • Spindle Apparatus / ultrastructure
  • Telophase / physiology
  • Time Factors
  • rho GTP-Binding Proteins / physiology

Substances

  • Benzenesulfonates
  • Cell Cycle Proteins
  • Etd1 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • C.I. Fluorescent Brightening Agent 28
  • Protein Kinases
  • Sid2 protein, S pombe
  • CDC2 Protein Kinase
  • rho GTP-Binding Proteins
  • rho1 protein, S pombe

Grant support

The research reported in this publication was supported by the grants BIO2015-69958-P to JCR and PP from the Ministerio de Economía y Competitividad (http://www.mineco.gob.es/) and CSI068P17 from MINECO/FEDER and Junta de Castilla y León (https://www.jcyl.es/) to PP and JCR. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.