TY - JOUR
T1 - Calcineurin-dependent Protein Phosphorylation Changes During Egg Activation in Drosophila melanogaster
AU - Zhang, Zijing
AU - Ahmed-Braimah, Yasir H.
AU - Goldberg, Michael L.
AU - Wolfner, Mariana F.
N1 - Funding Information:
Acknowledgments—We thank Dr. Tim Karr for the opportunity to contribute to this Special Issue, and NIH grant R21-HD088744 to M.F.W. for funding this study. We thank Drs. Terry Orr-Weaver (Massachusetts Institute of Technology) and Toshiro Aigaki for their kind gifts of anti-Gnu primary antibodies and CnAact flies, respectively. We thank Drs. Marcus Smolka (Cornell University), Steve Dorus (Syracuse University), Kathryn Lilley (University of Cambridge), and Sheng Zhang (Cornell BioResource Center) for helpful advice on the relative merits of different quantitative mass spectrometry methods, and Drs. John Schimenti and Ken Kemphues for helpful comments on an early version of this manuscript, and anonymous reviewers for helpful comments on this version. We thank the Proteomic and MS Facility of Cornell University’s BioResource Center for running the mass spectrometry on their Orbitrap Fusion mass spectrometer (which had been purchased with NIH grant SIG 1S10 OD017992). We thank the TRiP consortium at Harvard Medical School (funded by NIH grant NIGMS R01-GM084947) and the Bloomington Stock Center for transgenic RNAi fly stocks used in this study.
Funding Information:
We thank Dr. Tim Karr for the opportunity to contribute to this Special Issue, and NIH grant R21-HD088744 to M.F.W. for funding this study. We thank Drs. Terry Orr-Weaver (Massachusetts Institute of Technology) and Toshiro Aigaki for their kind gifts of anti-Gnu primary antibodies and CnAact flies, respectively. We thank Drs. Marcus Smolka (Cornell University), Steve Dorus (Syracuse University), Kathryn Lilley (University of Cambridge), and Sheng Zhang (Cornell BioResource Center) for helpful advice on the relative merits of different quantitative mass spectrometry methods, and Drs. John Schimenti and Ken Kemphues for helpful comments on an early version of this manuscript, and anonymous reviewers for helpful comments on this version. We thank the Proteomic and MS Facility of Cornell University’s BioResource Center for running the mass spectrometry on their Orbitrap Fusion mass spectrometer (which had been purchased with NIH grant SIG 1S10 OD017992). We thank the TRiP consortium at Harvard Medical School (funded by NIH grant NIGMS R01-GM084947) and the Bloomington Stock Center for transgenic RNAi fly stocks used in this study.
Publisher Copyright:
© 2019 Zhang et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2019/3
Y1 - 2019/3
N2 - In almost all animals studied to date, the crucial process of egg activation, by which an arrested mature oocyte transitions into an actively developing embryo, initiates with an increase in Ca 2 in the oocyte’s cytoplasm. This Ca 2 rise sets off a series of downstream events, including the completion of meiosis and the dynamic remodeling of the oocyte transcriptome and proteome, which prepares the oocyte for embryogenesis. Calcineurin is a highly conserved phosphatase that is activated by Ca 2 upon egg activation and that is required for the resumption of meiosis in Xenopus, ascidians, and Drosophila. The molecular mechanisms by which calcineurin transduces the calcium signal to regulate meiosis and other downstream events are still unclear. In this study, we investigate the regulatory role of calcineurin during egg activation in Drosophila melanogaster. Using mass spectrometry, we quantify the phosphoproteomic and proteomic changes that occur during egg activation, and we examine how these events are affected when calcineurin function is perturbed in female germ cells. Our results show that calcineurin regulates hundreds of phosphosites and also influences the abundance of numerous proteins during egg activation. We find calcineurin-dependent changes in cell cycle regulators including Fizzy (Fzy), Greatwall (Gwl) and Endosulfine (Endos); in protein translation modulators including PNG, NAT, eIF4G, and eIF4B; and in important components of signaling pathways including GSK3 and Akt1. Our results help elucidate the events that occur during the transition from oocyte to embryo.
AB - In almost all animals studied to date, the crucial process of egg activation, by which an arrested mature oocyte transitions into an actively developing embryo, initiates with an increase in Ca 2 in the oocyte’s cytoplasm. This Ca 2 rise sets off a series of downstream events, including the completion of meiosis and the dynamic remodeling of the oocyte transcriptome and proteome, which prepares the oocyte for embryogenesis. Calcineurin is a highly conserved phosphatase that is activated by Ca 2 upon egg activation and that is required for the resumption of meiosis in Xenopus, ascidians, and Drosophila. The molecular mechanisms by which calcineurin transduces the calcium signal to regulate meiosis and other downstream events are still unclear. In this study, we investigate the regulatory role of calcineurin during egg activation in Drosophila melanogaster. Using mass spectrometry, we quantify the phosphoproteomic and proteomic changes that occur during egg activation, and we examine how these events are affected when calcineurin function is perturbed in female germ cells. Our results show that calcineurin regulates hundreds of phosphosites and also influences the abundance of numerous proteins during egg activation. We find calcineurin-dependent changes in cell cycle regulators including Fizzy (Fzy), Greatwall (Gwl) and Endosulfine (Endos); in protein translation modulators including PNG, NAT, eIF4G, and eIF4B; and in important components of signaling pathways including GSK3 and Akt1. Our results help elucidate the events that occur during the transition from oocyte to embryo.
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U2 - 10.1074/mcp.RA118.001076
DO - 10.1074/mcp.RA118.001076
M3 - Article
C2 - 30478224
AN - SCOPUS:85063275936
SN - 1535-9476
VL - 18
SP - S145-S158
JO - Molecular and Cellular Proteomics
JF - Molecular and Cellular Proteomics
ER -