Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functions
Abstract RAD51C is a tumor suppressor gene with over 285 variants of unknown significance (VUS) found in primary ovarian tumors. RAD51C is a paralog of the recombinase RAD51, and it forms complexes with other paralogs to regulate RAD51 activity. We screened 27 ovarian cancer-derived RAD51C VUS to id...
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-61283-2 |
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| author | Hayley L. Rein Yashpal Rawal Anna L. Palovcak-Lightbourn Gayatri S. Ganesan Phoebe S. Parker Reagan Russell Kristie E. Darrah Mohammad Afsar Meghan R. Sullivan Sarah R. Hengel Marc R. Radke Patricia L. Opresko Judith L. Yanowitz Eric C. Greene Jung-Min Lee Susan M. Domchek Elizabeth M. Swisher Shaun K. Olsen Patrick Sung Kara A. Bernstein |
| author_facet | Hayley L. Rein Yashpal Rawal Anna L. Palovcak-Lightbourn Gayatri S. Ganesan Phoebe S. Parker Reagan Russell Kristie E. Darrah Mohammad Afsar Meghan R. Sullivan Sarah R. Hengel Marc R. Radke Patricia L. Opresko Judith L. Yanowitz Eric C. Greene Jung-Min Lee Susan M. Domchek Elizabeth M. Swisher Shaun K. Olsen Patrick Sung Kara A. Bernstein |
| author_sort | Hayley L. Rein |
| collection | DOAJ |
| description | Abstract RAD51C is a tumor suppressor gene with over 285 variants of unknown significance (VUS) found in primary ovarian tumors. RAD51C is a paralog of the recombinase RAD51, and it forms complexes with other paralogs to regulate RAD51 activity. We screened 27 ovarian cancer-derived RAD51C VUS to identify those that affect the assembly of functional tetrameric RAD51B-C-D-XRCC2 (BCDX2) complex. With yeast 3-hybrid and biochemical analyses, we identify a mutation cluster of the RAD51C Walker B region affecting protein interactions with other RAD51 paralogs. By further analyzing these variants for homologous recombination (HR), replication fork regression, DNA binding and ATPase activity, and RAD51 filament formation, we identified separation-of-function alleles that uncouple RAD51C distinct enzymatic activities with HR and replication. Thus, our analysis of RAD51C identifies additional VUS with functional defects, which will aid in pathogenicity classification and inform future strategies to treat individuals harboring RAD51C loss-of-function alleles. |
| format | Article |
| id | doaj-art-9dd67d52cc73427a9e3c04b41eb42da4 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-9dd67d52cc73427a9e3c04b41eb42da42025-08-20T03:42:52ZengNature PortfolioNature Communications2041-17232025-07-0116111310.1038/s41467-025-61283-2Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functionsHayley L. Rein0Yashpal Rawal1Anna L. Palovcak-Lightbourn2Gayatri S. Ganesan3Phoebe S. Parker4Reagan Russell5Kristie E. Darrah6Mohammad Afsar7Meghan R. Sullivan8Sarah R. Hengel9Marc R. Radke10Patricia L. Opresko11Judith L. Yanowitz12Eric C. Greene13Jung-Min Lee14Susan M. Domchek15Elizabeth M. Swisher16Shaun K. Olsen17Patrick Sung18Kara A. Bernstein19University of Pittsburgh, School of Medicine, Department of Pharmacology and Chemical BiologyUniversity of Texas Health Science Center at San Antonio, Department of Biochemistry and Structural Biology, Greehey Children’s Cancer Research InstituteUniversity of Pennsylvania School of Medicine, Department of Biochemistry and BiophysicsUniversity of Pennsylvania School of Medicine, Department of Biochemistry and BiophysicsUniversity of Pennsylvania School of Medicine, Department of Biochemistry and BiophysicsUniversity of Pittsburgh, School of Medicine, Department of Pharmacology and Chemical BiologyUniversity of Pennsylvania School of Medicine, Department of Biochemistry and BiophysicsUniversity of Texas Health Science Center at San Antonio, Department of Biochemistry and Structural Biology, Greehey Children’s Cancer Research InstituteUniversity of Pittsburgh, School of Medicine, Department of Microbiology and Molecular GeneticsUniversity of Pittsburgh, School of Medicine, Department of Pharmacology and Chemical BiologyDivision of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Washington School of MedicineDepartment of Environmental and Occupational Health, University of Pittsburgh School of Public HealthUniversity of Pittsburgh School of Medicine, Department of Obstetrics, Gynecology, and Reproductive ScienceDepartment of Biochemistry and Molecular Biophysics, Columbia University Medical CenterWomen’s Malignancies Branch, Center for Cancer Research, National Cancer InstituteBasser Center for BRCA, Abramson Cancer Centre, University of Pennsylvania School of MedicineDivision of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Washington School of MedicineUniversity of Texas Health Science Center at San Antonio, Department of Biochemistry and Structural Biology, Greehey Children’s Cancer Research InstituteUniversity of Texas Health Science Center at San Antonio, Department of Biochemistry and Structural Biology, Greehey Children’s Cancer Research InstituteUniversity of Pennsylvania School of Medicine, Department of Biochemistry and BiophysicsAbstract RAD51C is a tumor suppressor gene with over 285 variants of unknown significance (VUS) found in primary ovarian tumors. RAD51C is a paralog of the recombinase RAD51, and it forms complexes with other paralogs to regulate RAD51 activity. We screened 27 ovarian cancer-derived RAD51C VUS to identify those that affect the assembly of functional tetrameric RAD51B-C-D-XRCC2 (BCDX2) complex. With yeast 3-hybrid and biochemical analyses, we identify a mutation cluster of the RAD51C Walker B region affecting protein interactions with other RAD51 paralogs. By further analyzing these variants for homologous recombination (HR), replication fork regression, DNA binding and ATPase activity, and RAD51 filament formation, we identified separation-of-function alleles that uncouple RAD51C distinct enzymatic activities with HR and replication. Thus, our analysis of RAD51C identifies additional VUS with functional defects, which will aid in pathogenicity classification and inform future strategies to treat individuals harboring RAD51C loss-of-function alleles.https://doi.org/10.1038/s41467-025-61283-2 |
| spellingShingle | Hayley L. Rein Yashpal Rawal Anna L. Palovcak-Lightbourn Gayatri S. Ganesan Phoebe S. Parker Reagan Russell Kristie E. Darrah Mohammad Afsar Meghan R. Sullivan Sarah R. Hengel Marc R. Radke Patricia L. Opresko Judith L. Yanowitz Eric C. Greene Jung-Min Lee Susan M. Domchek Elizabeth M. Swisher Shaun K. Olsen Patrick Sung Kara A. Bernstein Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functions Nature Communications |
| title | Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functions |
| title_full | Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functions |
| title_fullStr | Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functions |
| title_full_unstemmed | Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functions |
| title_short | Comprehensive RAD51C ovarian cancer variant analysis uncouples homologous recombination and replicative functions |
| title_sort | comprehensive rad51c ovarian cancer variant analysis uncouples homologous recombination and replicative functions |
| url | https://doi.org/10.1038/s41467-025-61283-2 |
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