A large complex mediated by Moc1, Moc2 and Cpc2 regulates sexual differentiation in fission yeast.
Sexual differentiation in Schizosaccharomyces pombe is triggered by nutrient starvation and is downregulated by cAMP. Screening programs have identified the moc1/sds23, moc2/ded1, moc3 and moc4/zfs1 genes as inducers of sexual differentiation, even in the presence of elevated levels of cAMP. To investigate possible interactions among Moc1, Moc2, Moc3 and Moc4 proteins, we first screened for individual Moc-interacting proteins using the yeast two-hybrid system and verified the interactions with other Moc proteins. Using this screening process, Cpc2 and Rpl32-2 were highlighted as factors involved in interactions with multiple Moc proteins. Cpc2 interacted with Moc1, Moc2 and Moc3, whereas the ribosomal protein Rpl32-2 interacted with all Moc proteins in the two-hybrid system. Physical interactions of Cpc2 with Moc1, Moc2 and Rpl32-2, and of Rpl32-2 with Moc2 were confirmed by coimmunoprecipitation. In addition, using Blue Native/PAGE, we revealed that each Moc protein exists as a large complex. Overexpression of Moc1, Moc2, Moc3, Moc4 and Rpl32-2 resulted in the efficient induction of a key transcription factor Ste11, suggesting that all proteins tested are positive regulators of Ste11. Considering that Moc2/Ded1 is a general translation factor and that Cpc2 associates with many ribosomal proteins, including Rpl32-2, it is possible that a large Moc-mediated complex, detected in this study, may act as a translational regulator involved in the control of sexual differentiation in S. pombe through the induction of Ste11.
Paul
Swapan Kumar
SK
Department of Applied Bioscience and Biotechnology, Shimane University, Matsue, Japan.
Oowatari
Yasuo
Y
Kawamukai
Makoto
M
D018797:Cell Cycle Proteins / Q000502:physiology*; D053487:DEAD-box RNA Helicases / Q000502:physiology*; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000502:physiology*; D012568:Schizosaccharomyces / Q000502:physiology*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics; D020798:Two-Hybrid System Techniques
D018797:Cell Cycle Proteins; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D029702:Schizosaccharomyces pombe Proteins; D014157:Transcription Factors; C000617313:cpc2 protein, S pombe; C071596:ste11 protein, S pombe; C492151:sum3 protein, S pombe; D053487:DEAD-box RNA Helicases