Spo5 phosphorylation is essential for its own timely degradation and for successful meiosis in Schizosaccharomyces pombe.
Protein phosphorylation is pivotal for meiotic progression, but little is known about its regulatory mechanisms. We show that before meiosis I, the meiosis-specific Schizosaccharomyces pombe protein Spo5 is phosphorylated in vivo on T29, T55, S59 and/or T63. In a mutant strain expressing Spo5 fused to green fluorescent protein with alanine substitutions of these amino acid sites (GFP; Spo5-4A-GFP), the timely degradation of Spo5 at meiosis II was not observed. Additionally, Spo5-4A-GFP signals were retained after metaphase II and were localized to the nucleus. This was accompanied by the nuclear mislocalization of Psy1, a marker of the forespore membrane (FSM), and the generation of empty cells, in which cytoplasm had leaked from the ruptured membrane, as well as by the appearance of asci harboring deformed spores. Indeed, thin-section electron microscopy (TEM) revealed fragile-looking spo5-4A-GFP ascospores with ruffled spore walls. In contrast, a mutant strain expressing a constitutively-phosphorylated form of Spo5 (Spo5-4D-GFP) was phenotypically indistinguishable from a strain expressing wild-type (WT) protein (Spo5-WT-GFP). Taken together, these results indicate that Spo5 phosphorylation ensures the timely degradation of Spo5 during meiosis and the proper localization of Psy1, leading to the production of viable spores with robust FSMs and strong walls.
Okuzaki
Daisuke
D
Department of Molecular Genetics, Osaka University, Suita, Osaka, Japan.
Kasama
Takashi
T
Hirata
Aiko
A
Ohtaka
Ayami
A
Kakegawa
Reiko
R
Nojima
Hiroshi
H
D002467:Cell Nucleus / Q000378:metabolism; D008540:Meiosis; D008677:Metaphase; D009154:Mutation; D010766:Phosphorylation; D050765:Qa-SNARE Proteins / Q000032:analysis / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D012568:Schizosaccharomyces / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D013172:Spores, Fungal / Q000378:metabolism
C510389:Psy1 protein, S pombe; D050765:Qa-SNARE Proteins; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D029702:Schizosaccharomyces pombe Proteins; C516717:Spo5 protein, S pombe