Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes
Abstract FAM111A, a serine protease, plays roles in DNA replication and antiviral defense. Missense mutations in the catalytic domain cause hyper-autocleavage and are associated with genetic disorders with developmental defects. Despite the enzyme’s biological significance, the molecular architectur...
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| Format: | Article |
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Nature Portfolio
2024-03-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-46207-w |
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| author | Sowmiya Palani Yuka Machida Julia R. Alvey Vandana Mishra Allison L. Welter Gaofeng Cui Benoît Bragantini Maria Victoria Botuyan Anh T. Q. Cong Georges Mer Matthew J. Schellenberg Yuichi J. Machida |
| author_facet | Sowmiya Palani Yuka Machida Julia R. Alvey Vandana Mishra Allison L. Welter Gaofeng Cui Benoît Bragantini Maria Victoria Botuyan Anh T. Q. Cong Georges Mer Matthew J. Schellenberg Yuichi J. Machida |
| author_sort | Sowmiya Palani |
| collection | DOAJ |
| description | Abstract FAM111A, a serine protease, plays roles in DNA replication and antiviral defense. Missense mutations in the catalytic domain cause hyper-autocleavage and are associated with genetic disorders with developmental defects. Despite the enzyme’s biological significance, the molecular architecture of the FAM111A serine protease domain (SPD) is unknown. Here, we show that FAM111A is a dimerization-dependent protease containing a narrow, recessed active site that cleaves substrates with a chymotrypsin-like specificity. X-ray crystal structures and mutagenesis studies reveal that FAM111A dimerizes via the N-terminal helix within the SPD. This dimerization induces an activation cascade from the dimerization sensor loop to the oxyanion hole through disorder-to-order transitions. Dimerization is essential for proteolytic activity in vitro and for facilitating DNA replication at DNA-protein crosslink obstacles in cells, while it is dispensable for autocleavage. These findings underscore the role of dimerization in FAM111A’s function and highlight the distinction in its dimerization dependency between substrate cleavage and autocleavage. |
| format | Article |
| id | doaj-art-04b6d5d1449d4892b6e3d297c8272f1f |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2024-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-04b6d5d1449d4892b6e3d297c8272f1f2025-02-09T12:43:45ZengNature PortfolioNature Communications2041-17232024-03-0115111610.1038/s41467-024-46207-wDimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexesSowmiya Palani0Yuka Machida1Julia R. Alvey2Vandana Mishra3Allison L. Welter4Gaofeng Cui5Benoît Bragantini 6Maria Victoria Botuyan7Anh T. Q. Cong8Georges Mer9Matthew J. Schellenberg10Yuichi J. Machida11Mayo Clinic Graduate School of Biomedical Sciences, Mayo ClinicDepartment of Oncology, Division of Oncology Research, Mayo ClinicDepartment of Biochemistry and Molecular Biology, Mayo ClinicDevelopmental Therapeutics Branch, Center for Cancer Research, National Cancer InstituteMayo Clinic Graduate School of Biomedical Sciences, Mayo ClinicDepartment of Biochemistry and Molecular Biology, Mayo ClinicDepartment of Biochemistry and Molecular Biology, Mayo ClinicDepartment of Biochemistry and Molecular Biology, Mayo ClinicDepartment of Biochemistry and Molecular Biology, Mayo ClinicDepartment of Biochemistry and Molecular Biology, Mayo ClinicDepartment of Biochemistry and Molecular Biology, Mayo ClinicDepartment of Oncology, Division of Oncology Research, Mayo ClinicAbstract FAM111A, a serine protease, plays roles in DNA replication and antiviral defense. Missense mutations in the catalytic domain cause hyper-autocleavage and are associated with genetic disorders with developmental defects. Despite the enzyme’s biological significance, the molecular architecture of the FAM111A serine protease domain (SPD) is unknown. Here, we show that FAM111A is a dimerization-dependent protease containing a narrow, recessed active site that cleaves substrates with a chymotrypsin-like specificity. X-ray crystal structures and mutagenesis studies reveal that FAM111A dimerizes via the N-terminal helix within the SPD. This dimerization induces an activation cascade from the dimerization sensor loop to the oxyanion hole through disorder-to-order transitions. Dimerization is essential for proteolytic activity in vitro and for facilitating DNA replication at DNA-protein crosslink obstacles in cells, while it is dispensable for autocleavage. These findings underscore the role of dimerization in FAM111A’s function and highlight the distinction in its dimerization dependency between substrate cleavage and autocleavage.https://doi.org/10.1038/s41467-024-46207-w |
| spellingShingle | Sowmiya Palani Yuka Machida Julia R. Alvey Vandana Mishra Allison L. Welter Gaofeng Cui Benoît Bragantini Maria Victoria Botuyan Anh T. Q. Cong Georges Mer Matthew J. Schellenberg Yuichi J. Machida Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes Nature Communications |
| title | Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes |
| title_full | Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes |
| title_fullStr | Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes |
| title_full_unstemmed | Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes |
| title_short | Dimerization-dependent serine protease activity of FAM111A prevents replication fork stalling at topoisomerase 1 cleavage complexes |
| title_sort | dimerization dependent serine protease activity of fam111a prevents replication fork stalling at topoisomerase 1 cleavage complexes |
| url | https://doi.org/10.1038/s41467-024-46207-w |
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