A Huluwa phosphorylation switch regulates embryonic axis induction
Abstract Embryonic axis formation is essential for patterning and morphogenesis in vertebrates and is tightly regulated by the dorsal organizer. Previously, we demonstrated that maternally derived Huluwa (Hwa) acts as a dorsal determinant, dictating axis formation by activating β-catenin signaling i...
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Nature Portfolio
2024-11-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-54450-4 |
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| author | Yao Li Yun Yan Bo Gong Qianwen Zheng Haiyan Zhou Jiarui Sun Mingpeng Li Zhao Wang Yaohui Li Yunjing Wan Weixi Chen Shiqian Qi Xianming Mo Anming Meng Bo Xiang Jing Chen |
| author_facet | Yao Li Yun Yan Bo Gong Qianwen Zheng Haiyan Zhou Jiarui Sun Mingpeng Li Zhao Wang Yaohui Li Yunjing Wan Weixi Chen Shiqian Qi Xianming Mo Anming Meng Bo Xiang Jing Chen |
| author_sort | Yao Li |
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| description | Abstract Embryonic axis formation is essential for patterning and morphogenesis in vertebrates and is tightly regulated by the dorsal organizer. Previously, we demonstrated that maternally derived Huluwa (Hwa) acts as a dorsal determinant, dictating axis formation by activating β-catenin signaling in zebrafish and Xenopus. However, the mechanism of activation and fine regulation of the Hwa protein remains unclear. Through candidate screening we identified a mutation at Ser168 in the PPNSP motif of Hwa that dramatically abolishes its axis-inducing activity. Mechanistically, mutating the Ser168 residue reduced its binding affinity to Tankyrase 1/2 and the degradation of the Axin protein, weakening β-catenin signaling activation. We confirmed that Ser168 is phosphorylated and that phosphorylation increases Hwa activity in β-catenin signaling and axis induction. Several kinases including Cdk16, Cdk2, and GSK3β, were found to enhance Ser168 phosphorylation in vitro and in vivo. Both dominant-negative Cdk16 expression and pHwa (Ser168) antibody treatment reduce Hwa function. Lastly, a knock-in allele mutating Ser168 to alanine resulted in embryos lacking body axes, demonstrating that Ser168 is essential to axis formation. In summary, Ser168 acts as a phosphorylation switch in Hwa/β-catenin signaling for embryonic axis induction, regulated by multiple kinases. |
| format | Article |
| id | doaj-art-9e70f305737b42c49f08a1ae78992a8e |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Nature Portfolio |
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| series | Nature Communications |
| spelling | doaj-art-9e70f305737b42c49f08a1ae78992a8e2024-11-24T12:33:42ZengNature PortfolioNature Communications2041-17232024-11-0115111510.1038/s41467-024-54450-4A Huluwa phosphorylation switch regulates embryonic axis inductionYao Li0Yun Yan1Bo Gong2Qianwen Zheng3Haiyan Zhou4Jiarui Sun5Mingpeng Li6Zhao Wang7Yaohui Li8Yunjing Wan9Weixi Chen10Shiqian Qi11Xianming Mo12Anming Meng13Bo Xiang14Jing Chen15Department of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Cell and Developmental Biology, Weill Cornell Medicine, Cornell UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Urology, State Key Laboratory of Biotherapy, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Stem Cell Biology, State Key Laboratory of Biotherapy, West China Hospital, Sichuan UniversityLaboratory of Molecular Developmental Biology, State Key Laboratory of Membrane Biology, Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityDepartment of Pediatric Surgery and Laboratory of Pediatric Surgery, West China Hospital, Sichuan UniversityAbstract Embryonic axis formation is essential for patterning and morphogenesis in vertebrates and is tightly regulated by the dorsal organizer. Previously, we demonstrated that maternally derived Huluwa (Hwa) acts as a dorsal determinant, dictating axis formation by activating β-catenin signaling in zebrafish and Xenopus. However, the mechanism of activation and fine regulation of the Hwa protein remains unclear. Through candidate screening we identified a mutation at Ser168 in the PPNSP motif of Hwa that dramatically abolishes its axis-inducing activity. Mechanistically, mutating the Ser168 residue reduced its binding affinity to Tankyrase 1/2 and the degradation of the Axin protein, weakening β-catenin signaling activation. We confirmed that Ser168 is phosphorylated and that phosphorylation increases Hwa activity in β-catenin signaling and axis induction. Several kinases including Cdk16, Cdk2, and GSK3β, were found to enhance Ser168 phosphorylation in vitro and in vivo. Both dominant-negative Cdk16 expression and pHwa (Ser168) antibody treatment reduce Hwa function. Lastly, a knock-in allele mutating Ser168 to alanine resulted in embryos lacking body axes, demonstrating that Ser168 is essential to axis formation. In summary, Ser168 acts as a phosphorylation switch in Hwa/β-catenin signaling for embryonic axis induction, regulated by multiple kinases.https://doi.org/10.1038/s41467-024-54450-4 |
| spellingShingle | Yao Li Yun Yan Bo Gong Qianwen Zheng Haiyan Zhou Jiarui Sun Mingpeng Li Zhao Wang Yaohui Li Yunjing Wan Weixi Chen Shiqian Qi Xianming Mo Anming Meng Bo Xiang Jing Chen A Huluwa phosphorylation switch regulates embryonic axis induction Nature Communications |
| title | A Huluwa phosphorylation switch regulates embryonic axis induction |
| title_full | A Huluwa phosphorylation switch regulates embryonic axis induction |
| title_fullStr | A Huluwa phosphorylation switch regulates embryonic axis induction |
| title_full_unstemmed | A Huluwa phosphorylation switch regulates embryonic axis induction |
| title_short | A Huluwa phosphorylation switch regulates embryonic axis induction |
| title_sort | huluwa phosphorylation switch regulates embryonic axis induction |
| url | https://doi.org/10.1038/s41467-024-54450-4 |
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