Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i>
RAD51 is a central protein of homologous recombination and DNA repair processes that maintains genome stability and ensures the accurate repair of double-stranded breaks (DSBs). In this work, we assessed amyloid properties of RAD51 <i>in vitro</i> and in the bacterial curli-dependent amy...
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MDPI AG
2022-10-01
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| Series: | International Journal of Molecular Sciences |
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| author | Daniel V. Kachkin Kirill V. Volkov Julia V. Sopova Alexander G. Bobylev Sergei A. Fedotov Sergei G. Inge-Vechtomov Oxana V. Galzitskaya Yury O. Chernoff Aleksandr A. Rubel Anna Y. Aksenova |
| author_facet | Daniel V. Kachkin Kirill V. Volkov Julia V. Sopova Alexander G. Bobylev Sergei A. Fedotov Sergei G. Inge-Vechtomov Oxana V. Galzitskaya Yury O. Chernoff Aleksandr A. Rubel Anna Y. Aksenova |
| author_sort | Daniel V. Kachkin |
| collection | DOAJ |
| description | RAD51 is a central protein of homologous recombination and DNA repair processes that maintains genome stability and ensures the accurate repair of double-stranded breaks (DSBs). In this work, we assessed amyloid properties of RAD51 <i>in vitro</i> and in the bacterial curli-dependent amyloid generator (C-DAG) system. Resistance to ionic detergents, staining with amyloid-specific dyes, polarized microscopy, transmission electron microscopy (TEM), X-ray diffraction and other methods were used to evaluate the properties and structure of RAD51 aggregates. The purified human RAD51 protein formed detergent-resistant aggregates <i>in vitro</i> that had an unbranched cross-β fibrillar structure, which is typical for amyloids, and were stained with amyloid-specific dyes. Congo-red-stained RAD51 aggregates demonstrated birefringence under polarized light. RAD51 fibrils produced sharp circular X-ray reflections at 4.7 Å and 10 Å, demonstrating that they had a cross-β structure. Cytoplasmic aggregates of RAD51 were observed in cell cultures overexpressing <i>RAD51</i>. We demonstrated that a key protein that maintains genome stability, RAD51, has amyloid properties <i>in vitro</i> and in the C-DAG system and discussed the possible biological relevance of this observation. |
| first_indexed | 2024-03-09T21:38:17Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T21:38:17Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-7fefa267ef5c45c699b4d307962d020f2023-11-23T20:37:03ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-10-0123191165710.3390/ijms231911657Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i>Daniel V. Kachkin0Kirill V. Volkov1Julia V. Sopova2Alexander G. Bobylev3Sergei A. Fedotov4Sergei G. Inge-Vechtomov5Oxana V. Galzitskaya6Yury O. Chernoff7Aleksandr A. Rubel8Anna Y. Aksenova9Laboratory of Amyloid Biology, St. Petersburg State University, 199034 St. Petersburg, RussiaResearch Resource Center “Molecular and Cell Technologies”, Research Park, St. Petersburg State University (SPbSU), 199034 St. Petersburg, RussiaLaboratory of Amyloid Biology, St. Petersburg State University, 199034 St. Petersburg, RussiaInstitute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, 3 Institutskaya St., 142290 Moscow, RussiaLaboratory of Amyloid Biology, St. Petersburg State University, 199034 St. Petersburg, RussiaDepartment of Genetics and Biotechnology, St. Petersburg State University, 199034 St. Petersburg, RussiaInstitute of Theoretical and Experimental Biophysics, Russian Academy of Sciences, 3 Institutskaya St., 142290 Moscow, RussiaSchool of Biological Sciences, Georgia Institute of Technology, Atlanta, GA 30332-2000, USALaboratory of Amyloid Biology, St. Petersburg State University, 199034 St. Petersburg, RussiaLaboratory of Amyloid Biology, St. Petersburg State University, 199034 St. Petersburg, RussiaRAD51 is a central protein of homologous recombination and DNA repair processes that maintains genome stability and ensures the accurate repair of double-stranded breaks (DSBs). In this work, we assessed amyloid properties of RAD51 <i>in vitro</i> and in the bacterial curli-dependent amyloid generator (C-DAG) system. Resistance to ionic detergents, staining with amyloid-specific dyes, polarized microscopy, transmission electron microscopy (TEM), X-ray diffraction and other methods were used to evaluate the properties and structure of RAD51 aggregates. The purified human RAD51 protein formed detergent-resistant aggregates <i>in vitro</i> that had an unbranched cross-β fibrillar structure, which is typical for amyloids, and were stained with amyloid-specific dyes. Congo-red-stained RAD51 aggregates demonstrated birefringence under polarized light. RAD51 fibrils produced sharp circular X-ray reflections at 4.7 Å and 10 Å, demonstrating that they had a cross-β structure. Cytoplasmic aggregates of RAD51 were observed in cell cultures overexpressing <i>RAD51</i>. We demonstrated that a key protein that maintains genome stability, RAD51, has amyloid properties <i>in vitro</i> and in the C-DAG system and discussed the possible biological relevance of this observation.https://www.mdpi.com/1422-0067/23/19/11657RAD51protein aggregationprotein fibrilsamyloidX-ray diffractionfunctional amyloids |
| spellingShingle | Daniel V. Kachkin Kirill V. Volkov Julia V. Sopova Alexander G. Bobylev Sergei A. Fedotov Sergei G. Inge-Vechtomov Oxana V. Galzitskaya Yury O. Chernoff Aleksandr A. Rubel Anna Y. Aksenova Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i> International Journal of Molecular Sciences RAD51 protein aggregation protein fibrils amyloid X-ray diffraction functional amyloids |
| title | Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i> |
| title_full | Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i> |
| title_fullStr | Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i> |
| title_full_unstemmed | Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i> |
| title_short | Human RAD51 Protein Forms Amyloid-like Aggregates <i>In Vitro</i> |
| title_sort | human rad51 protein forms amyloid like aggregates i in vitro i |
| topic | RAD51 protein aggregation protein fibrils amyloid X-ray diffraction functional amyloids |
| url | https://www.mdpi.com/1422-0067/23/19/11657 |
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