Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803
Cyanobacteria, which have a photoautotrophic lifestyle, are threatened by ultraviolet solar rays and the reactive oxygen species generated during photosynthesis. They can adapt to environmental conditions primarily because of their DNA damage response and repair mechanisms, notably an efficient homo...
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| Language: | English |
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Frontiers Media S.A.
2024-04-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1362880/full |
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| author | Yanchao Gu Yantao Yang Chunhua Kou Ying Peng Wenguang Yang Jiayu Zhang Han Jin Xiaoru Han Yao Wang Xihui Shen |
| author_facet | Yanchao Gu Yantao Yang Chunhua Kou Ying Peng Wenguang Yang Jiayu Zhang Han Jin Xiaoru Han Yao Wang Xihui Shen |
| author_sort | Yanchao Gu |
| collection | DOAJ |
| description | Cyanobacteria, which have a photoautotrophic lifestyle, are threatened by ultraviolet solar rays and the reactive oxygen species generated during photosynthesis. They can adapt to environmental conditions primarily because of their DNA damage response and repair mechanisms, notably an efficient homologous recombination repair system. However, research on double-strand break (DSB) repair pathways, including the Holliday junction (HJ) resolution process, in Synechocystis sp. PCC6803 is limited. Here, we report that SynRuvC from cyanobacteria Synechocystis sp. PCC6803 has classical HJ resolution activity. We investigated the structural specificity, sequence preference, and biochemical properties of SynRuvC. SynRuvC strongly preferred Mn2+ as a cofactor, and its cleavage site predominantly resides within the 5′-TG↓(G/A)-3′ sequence. Interestingly, novel flap endonuclease and replication fork intermediate cleavage activities of SynRuvC were also determined, which distinguish it from other reported RuvCs. To explore the effect of SynRuvC on cell viability, we constructed a knockdown mutant and an overexpression strain of Synechocystis sp. PCC6803 (synruvCKD and synruvCOE) and assessed their survival under a variety of conditions. Knockdown of synruvC increased the sensitivity of cells to MMS, HU, and H2O2. The findings suggest that a novel RuvC family HJ resolvase SynRuvC is important in a variety of DNA repair processes and stress resistance in Synechocystis sp. PCC6803. |
| first_indexed | 2024-04-24T07:57:09Z |
| format | Article |
| id | doaj.art-68a092c5d7bb41fdbace69836bb5ad1c |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-04-24T07:57:09Z |
| publishDate | 2024-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-68a092c5d7bb41fdbace69836bb5ad1c2024-04-18T05:10:30ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-04-011510.3389/fmicb.2024.13628801362880Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803Yanchao Gu0Yantao Yang1Chunhua Kou2Ying Peng3Wenguang Yang4Jiayu Zhang5Han Jin6Xiaoru Han7Yao Wang8Xihui Shen9State Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaSuzhou XinBio Co., Ltd., Suzhou, Jiangsu, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaState Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Shaanxi Key Laboratory of Agricultural and Environmental Microbiology, College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, ChinaCyanobacteria, which have a photoautotrophic lifestyle, are threatened by ultraviolet solar rays and the reactive oxygen species generated during photosynthesis. They can adapt to environmental conditions primarily because of their DNA damage response and repair mechanisms, notably an efficient homologous recombination repair system. However, research on double-strand break (DSB) repair pathways, including the Holliday junction (HJ) resolution process, in Synechocystis sp. PCC6803 is limited. Here, we report that SynRuvC from cyanobacteria Synechocystis sp. PCC6803 has classical HJ resolution activity. We investigated the structural specificity, sequence preference, and biochemical properties of SynRuvC. SynRuvC strongly preferred Mn2+ as a cofactor, and its cleavage site predominantly resides within the 5′-TG↓(G/A)-3′ sequence. Interestingly, novel flap endonuclease and replication fork intermediate cleavage activities of SynRuvC were also determined, which distinguish it from other reported RuvCs. To explore the effect of SynRuvC on cell viability, we constructed a knockdown mutant and an overexpression strain of Synechocystis sp. PCC6803 (synruvCKD and synruvCOE) and assessed their survival under a variety of conditions. Knockdown of synruvC increased the sensitivity of cells to MMS, HU, and H2O2. The findings suggest that a novel RuvC family HJ resolvase SynRuvC is important in a variety of DNA repair processes and stress resistance in Synechocystis sp. PCC6803.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1362880/fullRuvCHolliday junction resolvasesflap endonuclease (FEN)replication fork intermediate (Ref-I) cleavage activitySynechocystis sp. PCC6803 |
| spellingShingle | Yanchao Gu Yantao Yang Chunhua Kou Ying Peng Wenguang Yang Jiayu Zhang Han Jin Xiaoru Han Yao Wang Xihui Shen Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803 Frontiers in Microbiology RuvC Holliday junction resolvases flap endonuclease (FEN) replication fork intermediate (Ref-I) cleavage activity Synechocystis sp. PCC6803 |
| title | Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803 |
| title_full | Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803 |
| title_fullStr | Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803 |
| title_full_unstemmed | Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803 |
| title_short | Classical and novel properties of Holliday junction resolvase SynRuvC from Synechocystis sp. PCC6803 |
| title_sort | classical and novel properties of holliday junction resolvase synruvc from synechocystis sp pcc6803 |
| topic | RuvC Holliday junction resolvases flap endonuclease (FEN) replication fork intermediate (Ref-I) cleavage activity Synechocystis sp. PCC6803 |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1362880/full |
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