title: Dephosphorylation of juxtamembrane serines and threonines of the NPR2 guanylyl cyclase is required for rapid resumption of oocyte meiosis in response to luteinizing hormone.
abstract: The meiotic cell cycle of mammalian oocytes starts during embryogenesis and then pauses until luteinizing hormone (LH) acts on the granulosa cells of the follicle surrounding the oocyte to restart the cell cycle. An essential event in this process is a decrease in cyclic GMP in the granulosa cells, and part of the cGMP decrease results from dephosphorylation and inactivation of the natriuretic peptide receptor 2 (NPR2) guanylyl cyclase, also known as guanylyl cyclase B. However, it is unknown whether NPR2 dephosphorylation is essential for LH-induced meiotic resumption. Here, we prevented NPR2 dephosphorylation by generating a mouse line in which the seven regulatory serines and threonines of NPR2 were changed to the phosphomimetic amino acid glutamate (Npr2-7E). Npr2-7E/7E follicles failed to show a decrease in enzyme activity in response to LH, and the cGMP decrease was attenuated; correspondingly, LH-induced meiotic resumption was delayed. Meiotic resumption in response to EGF receptor activation was likewise delayed, indicating that NPR2 dephosphorylation is a component of the pathway by which EGF receptor activation mediates LH signaling. We also found that most of the NPR2 protein in the follicle was present in the mural granulosa cells. These findings indicate that NPR2 dephosphorylation in the mural granulosa cells is essential for the normal progression of meiosis in response to LH and EGF receptor activation. In addition, these studies provide the first demonstration that a change in phosphorylation of a transmembrane guanylyl cyclase regulates a physiological process, a mechanism that may also control other developmental events.
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030, USA.
authors.forename: Leia C
authors.initials: LC
authors.lastname: Shuhaibar
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030, USA.
authors.forename: Jeremy R
authors.initials: JR
authors.lastname: Egbert
authors.affiliation: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Aaron B
authors.initials: AB
authors.lastname: Edmund
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030, USA.
authors.forename: Tracy F
authors.initials: TF
authors.lastname: Uliasz
authors.affiliation: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA.
authors.forename: Deborah M
authors.initials: DM
authors.lastname: Dickey
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030, USA.
authors.forename: Siu-Pok
authors.initials: SP
authors.lastname: Yee
authors.affiliation: Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN 55455, USA. Electronic address: potter@umn.edu.
authors.forename: Lincoln R
authors.initials: LR
authors.lastname: Potter
authors.affiliation: Department of Cell Biology, University of Connecticut Health Center, Farmington, CT 06030, USA. Electronic address: ljaffe@uchc.edu.
authors.forename: Laurinda A
authors.initials: LA
authors.lastname: Jaffe
mesh_terms: D000818:Animals; D006152:Cyclic GMP / Q000378:metabolism; D066260:Epiregulin / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism; D006162:Guanylate Cyclase / Q000378:metabolism; D007986:Luteinizing Hormone / Q000494:pharmacology*; D008540:Meiosis / Q000187:drug effects*; D051379:Mice; D009865:Oocytes / Q000166:cytology* / Q000201:enzymology*; D010766:Phosphorylation / Q000187:drug effects; D017461:Receptors, Atrial Natriuretic Factor / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*; D012756:Sheep; D013912:Threonine / Q000378:metabolism*; D000099242:Atrial Natriuretic Factor Receptor B
chemical_list: D006152:Cyclic GMP; D066260:Epiregulin; D006162:Guanylate Cyclase; D007986:Luteinizing Hormone; D017461:Receptors, Atrial Natriuretic Factor; D012694:Serine; D013912:Threonine; D000099242:Atrial Natriuretic Factor Receptor B