Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclases
The natriuretic peptide receptors NPR1 and NPR2, also known as guanylyl cyclase A and guanylyl cyclase B, have critical functions in many signaling pathways, but much remains unknown about their localization and function in vivo. To facilitate studies of these proteins, we developed genetically modi...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-10-01
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| Series: | Frontiers in Molecular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fnmol.2022.1007026/full |
| _version_ | 1811237585332207616 |
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| author | Jeremy R. Egbert Tracy F. Uliasz Katie M. Lowther Katie M. Lowther Deborah Kaback Deborah Kaback Brandon M. Wagner Chastity L. Healy Timothy D. O’Connell Lincoln R. Potter Lincoln R. Potter Laurinda A. Jaffe Siu-Pok Yee Siu-Pok Yee |
| author_facet | Jeremy R. Egbert Tracy F. Uliasz Katie M. Lowther Katie M. Lowther Deborah Kaback Deborah Kaback Brandon M. Wagner Chastity L. Healy Timothy D. O’Connell Lincoln R. Potter Lincoln R. Potter Laurinda A. Jaffe Siu-Pok Yee Siu-Pok Yee |
| author_sort | Jeremy R. Egbert |
| collection | DOAJ |
| description | The natriuretic peptide receptors NPR1 and NPR2, also known as guanylyl cyclase A and guanylyl cyclase B, have critical functions in many signaling pathways, but much remains unknown about their localization and function in vivo. To facilitate studies of these proteins, we developed genetically modified mouse lines in which endogenous NPR1 and NPR2 were tagged with the HA epitope. To investigate the role of phosphorylation in regulating NPR1 and NPR2 guanylyl cyclase activity, we developed mouse lines in which regulatory serines and threonines were substituted with glutamates, to mimic the negative charge of the phosphorylated forms (NPR1-8E and NPR2-7E). Here we describe the generation and applications of these mice. We show that the HA-NPR1 and HA-NPR2 mice can be used to characterize the relative expression levels of these proteins in different tissues. We describe studies using the NPR2-7E mice that indicate that dephosphorylation of NPR2 transduces signaling pathways in ovary and bone, and studies using the NPR1-8E mice that indicate that the phosphorylation state of NPR1 is a regulator of heart, testis, and adrenal function. |
| first_indexed | 2024-04-12T12:27:09Z |
| format | Article |
| id | doaj.art-03df6655e1294441a88babe71b7b9ad8 |
| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-04-12T12:27:09Z |
| publishDate | 2022-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-03df6655e1294441a88babe71b7b9ad82022-12-22T03:33:08ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992022-10-011510.3389/fnmol.2022.10070261007026Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclasesJeremy R. Egbert0Tracy F. Uliasz1Katie M. Lowther2Katie M. Lowther3Deborah Kaback4Deborah Kaback5Brandon M. Wagner6Chastity L. Healy7Timothy D. O’Connell8Lincoln R. Potter9Lincoln R. Potter10Laurinda A. Jaffe11Siu-Pok Yee12Siu-Pok Yee13Department of Cell Biology, University of Connecticut Health Center, Farmington, CT, United StatesDepartment of Cell Biology, University of Connecticut Health Center, Farmington, CT, United StatesDepartment of Cell Biology, University of Connecticut Health Center, Farmington, CT, United StatesCenter for Mouse Genome Modification, University of Connecticut Health Center, Farmington, CT, United StatesDepartment of Cell Biology, University of Connecticut Health Center, Farmington, CT, United StatesCenter for Mouse Genome Modification, University of Connecticut Health Center, Farmington, CT, United StatesDepartment of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, United StatesDepartment of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, United StatesDepartment of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, United StatesDepartment of Integrative Biology and Physiology, University of Minnesota, Minneapolis, MN, United StatesDepartment of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN, United StatesDepartment of Cell Biology, University of Connecticut Health Center, Farmington, CT, United StatesDepartment of Cell Biology, University of Connecticut Health Center, Farmington, CT, United StatesCenter for Mouse Genome Modification, University of Connecticut Health Center, Farmington, CT, United StatesThe natriuretic peptide receptors NPR1 and NPR2, also known as guanylyl cyclase A and guanylyl cyclase B, have critical functions in many signaling pathways, but much remains unknown about their localization and function in vivo. To facilitate studies of these proteins, we developed genetically modified mouse lines in which endogenous NPR1 and NPR2 were tagged with the HA epitope. To investigate the role of phosphorylation in regulating NPR1 and NPR2 guanylyl cyclase activity, we developed mouse lines in which regulatory serines and threonines were substituted with glutamates, to mimic the negative charge of the phosphorylated forms (NPR1-8E and NPR2-7E). Here we describe the generation and applications of these mice. We show that the HA-NPR1 and HA-NPR2 mice can be used to characterize the relative expression levels of these proteins in different tissues. We describe studies using the NPR2-7E mice that indicate that dephosphorylation of NPR2 transduces signaling pathways in ovary and bone, and studies using the NPR1-8E mice that indicate that the phosphorylation state of NPR1 is a regulator of heart, testis, and adrenal function.https://www.frontiersin.org/articles/10.3389/fnmol.2022.1007026/fullnatriuretic peptide receptorguanylyl cyclasephosphorylationgenetically modified micecyclic GMP |
| spellingShingle | Jeremy R. Egbert Tracy F. Uliasz Katie M. Lowther Katie M. Lowther Deborah Kaback Deborah Kaback Brandon M. Wagner Chastity L. Healy Timothy D. O’Connell Lincoln R. Potter Lincoln R. Potter Laurinda A. Jaffe Siu-Pok Yee Siu-Pok Yee Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclases Frontiers in Molecular Neuroscience natriuretic peptide receptor guanylyl cyclase phosphorylation genetically modified mice cyclic GMP |
| title | Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclases |
| title_full | Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclases |
| title_fullStr | Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclases |
| title_full_unstemmed | Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclases |
| title_short | Epitope-tagged and phosphomimetic mouse models for investigating natriuretic peptide-stimulated receptor guanylyl cyclases |
| title_sort | epitope tagged and phosphomimetic mouse models for investigating natriuretic peptide stimulated receptor guanylyl cyclases |
| topic | natriuretic peptide receptor guanylyl cyclase phosphorylation genetically modified mice cyclic GMP |
| url | https://www.frontiersin.org/articles/10.3389/fnmol.2022.1007026/full |
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