The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i>
The study of DNA repair in hyperthermophiles has the potential to elucidate the mechanisms of genome integrity maintenance systems under extreme conditions. Previous biochemical studies have suggested that the single-stranded DNA-binding protein (SSB) from the hyperthermophilic crenarchaeon <i>...
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MDPI AG
2023-02-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/5/4558 |
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| author | Shoji Suzuki Norio Kurosawa |
| author_facet | Shoji Suzuki Norio Kurosawa |
| author_sort | Shoji Suzuki |
| collection | DOAJ |
| description | The study of DNA repair in hyperthermophiles has the potential to elucidate the mechanisms of genome integrity maintenance systems under extreme conditions. Previous biochemical studies have suggested that the single-stranded DNA-binding protein (SSB) from the hyperthermophilic crenarchaeon <i>Sulfolobus</i> is involved in the maintenance of genome integrity, namely, in mutation avoidance, homologous recombination (HR), and the repair of helix-distorting DNA lesions. However, no genetic study has been reported that elucidates whether SSB actually maintains genome integrity in <i>Sulfolobus</i> in vivo. Here, we characterized mutant phenotypes of the <i>ssb</i>-deleted strain Δ<i>ssb</i> in the thermophilic crenarchaeon <i>S. acidocaldarius</i>. Notably, an increase (29-fold) in mutation rate and a defect in HR frequency was observed in Δ<i>ssb</i>, indicating that SSB was involved in mutation avoidance and HR in vivo. We characterized the sensitivities of Δ<i>ssb</i>, in parallel with putative SSB-interacting protein-encoding gene-deleted strains, to DNA-damaging agents. The results showed that not only Δ<i>ssb</i> but also Δ<i>alhr1</i> and ΔSaci_0790 were markedly sensitive to a wide variety of helix-distorting DNA-damaging agents, indicating that SSB, a novel helicase <i>Sac</i>aLhr1, and a hypothetical protein Saci_0790, were involved in the repair of helix-distorting DNA lesions. This study expands our knowledge of the impact of SSB on genome integrity and identifies novel and key proteins for genome integrity in hyperthermophilic archaea in vivo. |
| first_indexed | 2024-03-11T07:22:50Z |
| format | Article |
| id | doaj.art-9ffe94b82faa4443ad5dcb9c502cc660 |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T07:22:50Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-9ffe94b82faa4443ad5dcb9c502cc6602023-11-17T07:50:09ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-02-01245455810.3390/ijms24054558The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i>Shoji Suzuki0Norio Kurosawa1Super-Cutting-Edge Grand and Advanced Research (SUGAR) Program, Institute for Extra-Cutting-Edge Science and Technology Avant-Garde Research (X-Star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka 237-0061, JapanDepartment of Science and Engineering for Sustainable Innovation, Faculty of Science and Engineering, Soka University, Hachioji 192-8577, JapanThe study of DNA repair in hyperthermophiles has the potential to elucidate the mechanisms of genome integrity maintenance systems under extreme conditions. Previous biochemical studies have suggested that the single-stranded DNA-binding protein (SSB) from the hyperthermophilic crenarchaeon <i>Sulfolobus</i> is involved in the maintenance of genome integrity, namely, in mutation avoidance, homologous recombination (HR), and the repair of helix-distorting DNA lesions. However, no genetic study has been reported that elucidates whether SSB actually maintains genome integrity in <i>Sulfolobus</i> in vivo. Here, we characterized mutant phenotypes of the <i>ssb</i>-deleted strain Δ<i>ssb</i> in the thermophilic crenarchaeon <i>S. acidocaldarius</i>. Notably, an increase (29-fold) in mutation rate and a defect in HR frequency was observed in Δ<i>ssb</i>, indicating that SSB was involved in mutation avoidance and HR in vivo. We characterized the sensitivities of Δ<i>ssb</i>, in parallel with putative SSB-interacting protein-encoding gene-deleted strains, to DNA-damaging agents. The results showed that not only Δ<i>ssb</i> but also Δ<i>alhr1</i> and ΔSaci_0790 were markedly sensitive to a wide variety of helix-distorting DNA-damaging agents, indicating that SSB, a novel helicase <i>Sac</i>aLhr1, and a hypothetical protein Saci_0790, were involved in the repair of helix-distorting DNA lesions. This study expands our knowledge of the impact of SSB on genome integrity and identifies novel and key proteins for genome integrity in hyperthermophilic archaea in vivo.https://www.mdpi.com/1422-0067/24/5/4558hyperthermophilic archaeamutation avoidancehomologous recombinationDNA repair<i>Sulfolobus acidocaldarius</i> |
| spellingShingle | Shoji Suzuki Norio Kurosawa The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i> International Journal of Molecular Sciences hyperthermophilic archaea mutation avoidance homologous recombination DNA repair <i>Sulfolobus acidocaldarius</i> |
| title | The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i> |
| title_full | The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i> |
| title_fullStr | The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i> |
| title_full_unstemmed | The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i> |
| title_short | The Impact of Single-Stranded DNA-Binding Protein SSB and Putative SSB-Interacting Proteins on Genome Integrity in the Thermophilic Crenarchaeon <i>Sulfolobus acidocaldarius</i> |
| title_sort | impact of single stranded dna binding protein ssb and putative ssb interacting proteins on genome integrity in the thermophilic crenarchaeon i sulfolobus acidocaldarius i |
| topic | hyperthermophilic archaea mutation avoidance homologous recombination DNA repair <i>Sulfolobus acidocaldarius</i> |
| url | https://www.mdpi.com/1422-0067/24/5/4558 |
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