Abstract

In Schizosaccharomyces pombe, the spore wall confers strong resistance against external stress. During meiosis II, the double-layered intracellular forespore membrane (FSM) forms de novo and encapsulates the nucleus. Eventually, the inner FSM layer becomes the plasma membrane of the spore, while the outer layer breaks down. However, the molecular mechanism and biological significance of this membrane breakdown remain unknown. Here, by genetic investigation of an S. pombe mutant (E22) with normal prespore formation but abnormal spores, we showed that Meu5, an RNA-binding protein known to bind to and stabilize more than 80 transcripts, is involved in this process. We confirmed that the E22 mutant does not produce Meu5 protein, while overexpression of meu5+ in E22 restores the sporulation defect. Furthermore, electron microscopy revealed that the outer membrane of the FSM persisted in meu5∆ spores. Investigation of the target genes of meu5+ showed that a mutant of cyc1+ encoding cytochrome c also showed a severe defect in outer FSM breakdown. Lastly, we determined that outer FSM breakdown occurs coincident with or after formation of the outermost Isp3 layer of the spore wall. Collectively, our data provide novel insights into the molecular mechanism of spore formation.

Highlights

  • Sporulation in the fission yeast Schizosaccharomyces pombe is a common cellular process that occurs in response to nutrition starvation, especially nitrogen starvation [1,2]

  • In S. pombe and the prospore membrane in S. cerevisiae) comprises two units, and the spore wall is formed between these two units

  • Electron microscopy has revealed that, in S. pombe, the outer forespore membrane (FSM) disappears during spore wall formation [1]

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Summary

Introduction

Sporulation in the fission yeast Schizosaccharomyces pombe is a common cellular process that occurs in response to nutrition starvation, especially nitrogen starvation [1,2]. The most significant difference in the spore wall structure between these two yeasts is the outermost layer: in S. cerevisiae, the outermost layer is composed mainly of dityrosine [12], whereas the S. pombe spore is covered by an extraordinarily abundant protein termed Isp3 [18,19]. In both yeasts, the outer layer of the FSM (corresponding to the prospore membrane in S. cerevisiae) dissolves during sporulation [1,20], suggesting that breakdown of this membrane contributes to proper spore wall formation. We propose the possibility that breakdown of the S. pombe outer FSM occurs after or coincident with the formation of the outermost spore layer of Isp

Materials and Methods
Fluorescence Microscopy
Quick-Freeze Deep-Etch Electron Microscopy
Thin-Section Electron Microscopy
Reverse Transcription PCR
Western Blotting
The E22 Mutant a Defect in Spore Maturation
Normal
Meu5 Is Involved in Breakdown of the Outer FSM
Quick-freeze
Cytochrome c IscInvolved inin
Localization of Isp3 to the SporeofPeriphery
Discussion
Methods
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