FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination
Background An impeccable female meiotic prophase is critical for producing a high‐quality oocyte and, ultimately, a healthy newborn. SYCP3 is a key component of the synaptonemal complex regulating meiotic homologous recombination. However, what regulates SYCP3 stability is unknown. Methods Fertility...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2022-07-01
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| Series: | Clinical and Translational Medicine |
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| Online Access: | https://doi.org/10.1002/ctm2.891 |
| _version_ | 1811221635974299648 |
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| author | Yang Wang Wen‐Yi Gao Li‐Li Wang Ruo‐Lei Wang Zhi‐Xia Yang Fu‐Qiang Luo Yu‐Hao He Zi‐Bin Wang Fu‐Qiang Wang Qing‐Yuan Sun Jing Li Dong Zhang |
| author_facet | Yang Wang Wen‐Yi Gao Li‐Li Wang Ruo‐Lei Wang Zhi‐Xia Yang Fu‐Qiang Luo Yu‐Hao He Zi‐Bin Wang Fu‐Qiang Wang Qing‐Yuan Sun Jing Li Dong Zhang |
| author_sort | Yang Wang |
| collection | DOAJ |
| description | Background An impeccable female meiotic prophase is critical for producing a high‐quality oocyte and, ultimately, a healthy newborn. SYCP3 is a key component of the synaptonemal complex regulating meiotic homologous recombination. However, what regulates SYCP3 stability is unknown. Methods Fertility assays, follicle counting, meiotic prophase stage (leptotene, zygotene, pachytene and diplotene) analysis and live imaging were employed to examine how FBXW24 knockout (KO) affect female fertility, follicle reserve, oocyte quality, meiotic prophase progression of female germ cells, and meiosis of oocytes. Western blot and immunostaining were used to examined the levels & signals (intensity, foci) of SYCP3 and multiple key DSB indicators & repair proteins (γH2AX, RPA2, p‐CHK2, RAD51, MLH1, HORMAD1, TRIP13) after FBXW24 KO. Co‐IP and immuno‐EM were used to examined the interaction between FBXW24 and SYCP3; Mass spec was used to characterize the ubiquitination sites in SYCP3; In vivo & in vitro ubiquitination assays were utilized to determine the key sites in SYCP3 & FBXW24 for ubiquitination. Results Fbxw24‐knockout (KO) female mice were infertile due to massive oocyte death upon meiosis entry. Fbxw24‐KO oocytes were defective due to elevated DNA double‐strand breaks (DSBs) and inseparable homologous chromosomes. Fbxw24‐KO germ cells showed increased SYCP3 levels, delayed prophase progression, increased DSBs, and decreased crossover foci. Next, we found that FBXW24 directly binds and ubiquitinates SYCP3 to regulate its stability. In addition, several key residues important for SYCP3 ubiquitination and FBXW24 ubiquitinating activity were characterized. Conclusions We proposed that FBXW24 regulates the timely degradation of SYCP3 to ensure normal crossover and DSB repair during pachytene. FBXW24‐KO delayed SYCP3 degradation and DSB repair from pachytene until metaphase II (MII), ultimately causing failure in oocyte maturation, oocyte death, and infertility. |
| first_indexed | 2024-04-12T08:04:04Z |
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| id | doaj.art-09f7e17f62a04f0a8aabce007e4effdc |
| institution | Directory Open Access Journal |
| issn | 2001-1326 |
| language | English |
| last_indexed | 2024-04-12T08:04:04Z |
| publishDate | 2022-07-01 |
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| series | Clinical and Translational Medicine |
| spelling | doaj.art-09f7e17f62a04f0a8aabce007e4effdc2022-12-22T03:41:14ZengWileyClinical and Translational Medicine2001-13262022-07-01127n/an/a10.1002/ctm2.891FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitinationYang Wang0Wen‐Yi Gao1Li‐Li Wang2Ruo‐Lei Wang3Zhi‐Xia Yang4Fu‐Qiang Luo5Yu‐Hao He6Zi‐Bin Wang7Fu‐Qiang Wang8Qing‐Yuan Sun9Jing Li10Dong Zhang11State Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaAnalysis and Test Center Nanjing Medical University Nanjing ChinaFertility Preservation Lab and Guangdong‐Hong Kong Metabolism & Reproduction Joint Laboratory Reproductive Medicine Center Guangdong Second Provincial General Hospital Guangzhou ChinaFertility Preservation Lab and Guangdong‐Hong Kong Metabolism & Reproduction Joint Laboratory Reproductive Medicine Center Guangdong Second Provincial General Hospital Guangzhou ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaState Key Lab of Reproductive Medicine Nanjing Medical University Nanjing ChinaBackground An impeccable female meiotic prophase is critical for producing a high‐quality oocyte and, ultimately, a healthy newborn. SYCP3 is a key component of the synaptonemal complex regulating meiotic homologous recombination. However, what regulates SYCP3 stability is unknown. Methods Fertility assays, follicle counting, meiotic prophase stage (leptotene, zygotene, pachytene and diplotene) analysis and live imaging were employed to examine how FBXW24 knockout (KO) affect female fertility, follicle reserve, oocyte quality, meiotic prophase progression of female germ cells, and meiosis of oocytes. Western blot and immunostaining were used to examined the levels & signals (intensity, foci) of SYCP3 and multiple key DSB indicators & repair proteins (γH2AX, RPA2, p‐CHK2, RAD51, MLH1, HORMAD1, TRIP13) after FBXW24 KO. Co‐IP and immuno‐EM were used to examined the interaction between FBXW24 and SYCP3; Mass spec was used to characterize the ubiquitination sites in SYCP3; In vivo & in vitro ubiquitination assays were utilized to determine the key sites in SYCP3 & FBXW24 for ubiquitination. Results Fbxw24‐knockout (KO) female mice were infertile due to massive oocyte death upon meiosis entry. Fbxw24‐KO oocytes were defective due to elevated DNA double‐strand breaks (DSBs) and inseparable homologous chromosomes. Fbxw24‐KO germ cells showed increased SYCP3 levels, delayed prophase progression, increased DSBs, and decreased crossover foci. Next, we found that FBXW24 directly binds and ubiquitinates SYCP3 to regulate its stability. In addition, several key residues important for SYCP3 ubiquitination and FBXW24 ubiquitinating activity were characterized. Conclusions We proposed that FBXW24 regulates the timely degradation of SYCP3 to ensure normal crossover and DSB repair during pachytene. FBXW24‐KO delayed SYCP3 degradation and DSB repair from pachytene until metaphase II (MII), ultimately causing failure in oocyte maturation, oocyte death, and infertility.https://doi.org/10.1002/ctm2.891FBXW24meiotic prophaseoocyteSYCP3ubiquitination |
| spellingShingle | Yang Wang Wen‐Yi Gao Li‐Li Wang Ruo‐Lei Wang Zhi‐Xia Yang Fu‐Qiang Luo Yu‐Hao He Zi‐Bin Wang Fu‐Qiang Wang Qing‐Yuan Sun Jing Li Dong Zhang FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination Clinical and Translational Medicine FBXW24 meiotic prophase oocyte SYCP3 ubiquitination |
| title | FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination |
| title_full | FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination |
| title_fullStr | FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination |
| title_full_unstemmed | FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination |
| title_short | FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination |
| title_sort | fbxw24 controls female meiotic prophase progression by regulating sycp3 ubiquitination |
| topic | FBXW24 meiotic prophase oocyte SYCP3 ubiquitination |
| url | https://doi.org/10.1002/ctm2.891 |
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