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Phosphoproteomics of ATR signaling in mouse testes
The phosphatidylinositol 3′ kinase (PI3K)‐related kinase ATR is crucial for mammalian meiosis. ATR promotes meiotic progression by coordinating key events in DNA repair, meiotic sex chromosome inactivation (MSCI), and checkpoint-dependent quality control during meiotic prophase I. Despite its centra...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8824463/ https://www.ncbi.nlm.nih.gov/pubmed/35133275 http://dx.doi.org/10.7554/eLife.68648 |
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author | Sims, Jennie R Faça, Vitor M Pereira, Catalina Ascenção, Carolline Comstock, William Badar, Jumana Arroyo-Martinez, Gerardo A Freire, Raimundo Cohen, Paula E Weiss, Robert S Smolka, Marcus B |
author_facet | Sims, Jennie R Faça, Vitor M Pereira, Catalina Ascenção, Carolline Comstock, William Badar, Jumana Arroyo-Martinez, Gerardo A Freire, Raimundo Cohen, Paula E Weiss, Robert S Smolka, Marcus B |
author_sort | Sims, Jennie R |
collection | PubMed |
description | The phosphatidylinositol 3′ kinase (PI3K)‐related kinase ATR is crucial for mammalian meiosis. ATR promotes meiotic progression by coordinating key events in DNA repair, meiotic sex chromosome inactivation (MSCI), and checkpoint-dependent quality control during meiotic prophase I. Despite its central roles in meiosis, the ATR-dependent meiotic signaling network remains largely unknown. Here, we used phosphoproteomics to define ATR signaling events in testes from mice following chemical and genetic ablation of ATR signaling. Quantitative analysis of phosphoproteomes obtained after germ cell-specific genetic ablation of the ATR activating 9-1-1 complex or treatment with ATR inhibitor identified over 14,000 phosphorylation sites from testes samples, of which 401 phosphorylation sites were found to be dependent on both the 9-1-1 complex and ATR. Our analyses identified ATR-dependent phosphorylation events in crucial DNA damage signaling and DNA repair proteins including TOPBP1, SMC3, MDC1, RAD50, and SLX4. Importantly, we identified ATR and RAD1-dependent phosphorylation events in proteins involved in mRNA regulatory processes, including SETX and RANBP3, whose localization to the sex body was lost upon ATR inhibition. In addition to identifying the expected ATR-targeted S/T-Q motif, we identified enrichment of an S/T-P-X-K motif in the set of ATR-dependent events, suggesting that ATR promotes signaling via proline-directed kinase(s) during meiosis. Indeed, we found that ATR signaling is important for the proper localization of CDK2 in spermatocytes. Overall, our analysis establishes a map of ATR signaling in mouse testes and highlights potential meiotic-specific actions of ATR during prophase I progression. |
format | Online Article Text |
id | pubmed-8824463 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-88244632022-02-10 Phosphoproteomics of ATR signaling in mouse testes Sims, Jennie R Faça, Vitor M Pereira, Catalina Ascenção, Carolline Comstock, William Badar, Jumana Arroyo-Martinez, Gerardo A Freire, Raimundo Cohen, Paula E Weiss, Robert S Smolka, Marcus B eLife Biochemistry and Chemical Biology The phosphatidylinositol 3′ kinase (PI3K)‐related kinase ATR is crucial for mammalian meiosis. ATR promotes meiotic progression by coordinating key events in DNA repair, meiotic sex chromosome inactivation (MSCI), and checkpoint-dependent quality control during meiotic prophase I. Despite its central roles in meiosis, the ATR-dependent meiotic signaling network remains largely unknown. Here, we used phosphoproteomics to define ATR signaling events in testes from mice following chemical and genetic ablation of ATR signaling. Quantitative analysis of phosphoproteomes obtained after germ cell-specific genetic ablation of the ATR activating 9-1-1 complex or treatment with ATR inhibitor identified over 14,000 phosphorylation sites from testes samples, of which 401 phosphorylation sites were found to be dependent on both the 9-1-1 complex and ATR. Our analyses identified ATR-dependent phosphorylation events in crucial DNA damage signaling and DNA repair proteins including TOPBP1, SMC3, MDC1, RAD50, and SLX4. Importantly, we identified ATR and RAD1-dependent phosphorylation events in proteins involved in mRNA regulatory processes, including SETX and RANBP3, whose localization to the sex body was lost upon ATR inhibition. In addition to identifying the expected ATR-targeted S/T-Q motif, we identified enrichment of an S/T-P-X-K motif in the set of ATR-dependent events, suggesting that ATR promotes signaling via proline-directed kinase(s) during meiosis. Indeed, we found that ATR signaling is important for the proper localization of CDK2 in spermatocytes. Overall, our analysis establishes a map of ATR signaling in mouse testes and highlights potential meiotic-specific actions of ATR during prophase I progression. eLife Sciences Publications, Ltd 2022-02-08 /pmc/articles/PMC8824463/ /pubmed/35133275 http://dx.doi.org/10.7554/eLife.68648 Text en © 2022, Sims et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Biochemistry and Chemical Biology Sims, Jennie R Faça, Vitor M Pereira, Catalina Ascenção, Carolline Comstock, William Badar, Jumana Arroyo-Martinez, Gerardo A Freire, Raimundo Cohen, Paula E Weiss, Robert S Smolka, Marcus B Phosphoproteomics of ATR signaling in mouse testes |
title | Phosphoproteomics of ATR signaling in mouse testes |
title_full | Phosphoproteomics of ATR signaling in mouse testes |
title_fullStr | Phosphoproteomics of ATR signaling in mouse testes |
title_full_unstemmed | Phosphoproteomics of ATR signaling in mouse testes |
title_short | Phosphoproteomics of ATR signaling in mouse testes |
title_sort | phosphoproteomics of atr signaling in mouse testes |
topic | Biochemistry and Chemical Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8824463/ https://www.ncbi.nlm.nih.gov/pubmed/35133275 http://dx.doi.org/10.7554/eLife.68648 |
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