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

Juan Carlos G Cortés,Iris Barragán,Pilar Pérez,Sergio Moreno,Juan Carlos Ribas,Masako Osumi,María Alcaide-Gavilán,Mami Konomi,M Belén Moreno,Mariona Ramos,Dannel Mccollum

doi:10.1371/journal.pgen.1007388

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.

Highlights

The division of a cell into two genetically identical daughter cells requires the accurate coordination of events such as the entry into mitosis, chromosome segregation, and cytokinesis
Fission yeast cytokinesis requires the invagination of the equatorial plasma membrane coupled to the synthesis of a special wall structure named septum
Despite Cdk1 kinase is inactivated in early anaphase, it is believed that cleavage furrow ingression and septation onset require anaphase progression and mitosis completion, only initiating after the complete activation of the Hippo-like septation initiation network (SIN) after telophase onset

Summary

Introduction

The division of a cell into two genetically identical daughter cells requires the accurate coordination of events such as the entry into mitosis, chromosome segregation, and cytokinesis. At the end of anaphase B the chromosomes decondense, the spindle breaks down, and cleavage furrow ingression begins [1,2,3]. Progression through the cell cycle is controlled by fluctuations in the activity of cyclin-dependent kinase (Cdk) complexes, which are dependent upon periodic expression of cyclin subunits. At anaphase B onset, cyclin is degraded by the anaphase-promoting complex which allows anaphase B progression and the exit from mitosis [4]. The expression of a non-degradable cyclin B in fission yeast blocks the cells in anaphase B and abolishes cytokinesis [7]

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Discussion

Conclusion