An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA binding
Abstract AcrIIA15 is an anti-CRISPR (Acr) protein that inhibits Staphylococcus aureus Cas9 (SaCas9). Although previous studies suggested it has dual functions, the structural and biochemical basis for its two activities remains unclear. Here, we determined the cryo-EM structure of AcrIIA15 in comple...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2024-02-01
|
| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-45987-5 |
| _version_ | 1797273969000185856 |
|---|---|
| author | Xieshuting Deng Wei Sun Xueyan Li Jiuyu Wang Zhi Cheng Gang Sheng Yanli Wang |
| author_facet | Xieshuting Deng Wei Sun Xueyan Li Jiuyu Wang Zhi Cheng Gang Sheng Yanli Wang |
| author_sort | Xieshuting Deng |
| collection | DOAJ |
| description | Abstract AcrIIA15 is an anti-CRISPR (Acr) protein that inhibits Staphylococcus aureus Cas9 (SaCas9). Although previous studies suggested it has dual functions, the structural and biochemical basis for its two activities remains unclear. Here, we determined the cryo-EM structure of AcrIIA15 in complex with SaCas9-sgRNA to reveal the inhibitory mechanism of the Acr’s C-terminal domain (CTD) in mimicking dsDNA to block protospacer adjacent motif (PAM) recognition. For the N-terminal domain (NTD), our crystal structures of the AcrIIA15-promoter DNA show that AcrIIA15 dimerizes through its NTD to recognize double-stranded (ds) DNA. Further, AcrIIA15 can simultaneously bind to both SaCas9-sgRNA and promoter DNA, creating a supercomplex of two Cas9s bound to two CTDs converging on a dimer of the NTD bound to a dsDNA. These findings shed light on AcrIIA15’s inhibitory mechanisms and its autoregulation of transcription, enhancing our understanding of phage-host interactions and CRISPR defense. |
| first_indexed | 2024-03-07T14:51:46Z |
| format | Article |
| id | doaj.art-9751cd020e0e4484a5aec6e1e7391be4 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-07T14:51:46Z |
| publishDate | 2024-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-9751cd020e0e4484a5aec6e1e7391be42024-03-05T19:41:22ZengNature PortfolioNature Communications2041-17232024-02-0115111310.1038/s41467-024-45987-5An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA bindingXieshuting Deng0Wei Sun1Xueyan Li2Jiuyu Wang3Zhi Cheng4Gang Sheng5Yanli Wang6Key Laboratory of RNA Science and Engineering, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of RNA Science and Engineering, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of RNA Science and Engineering, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of RNA Science and Engineering, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of RNA Science and Engineering, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of RNA Science and Engineering, Institute of Biophysics, Chinese Academy of SciencesKey Laboratory of RNA Science and Engineering, Institute of Biophysics, Chinese Academy of SciencesAbstract AcrIIA15 is an anti-CRISPR (Acr) protein that inhibits Staphylococcus aureus Cas9 (SaCas9). Although previous studies suggested it has dual functions, the structural and biochemical basis for its two activities remains unclear. Here, we determined the cryo-EM structure of AcrIIA15 in complex with SaCas9-sgRNA to reveal the inhibitory mechanism of the Acr’s C-terminal domain (CTD) in mimicking dsDNA to block protospacer adjacent motif (PAM) recognition. For the N-terminal domain (NTD), our crystal structures of the AcrIIA15-promoter DNA show that AcrIIA15 dimerizes through its NTD to recognize double-stranded (ds) DNA. Further, AcrIIA15 can simultaneously bind to both SaCas9-sgRNA and promoter DNA, creating a supercomplex of two Cas9s bound to two CTDs converging on a dimer of the NTD bound to a dsDNA. These findings shed light on AcrIIA15’s inhibitory mechanisms and its autoregulation of transcription, enhancing our understanding of phage-host interactions and CRISPR defense.https://doi.org/10.1038/s41467-024-45987-5 |
| spellingShingle | Xieshuting Deng Wei Sun Xueyan Li Jiuyu Wang Zhi Cheng Gang Sheng Yanli Wang An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA binding Nature Communications |
| title | An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA binding |
| title_full | An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA binding |
| title_fullStr | An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA binding |
| title_full_unstemmed | An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA binding |
| title_short | An anti-CRISPR that represses its own transcription while blocking Cas9-target DNA binding |
| title_sort | anti crispr that represses its own transcription while blocking cas9 target dna binding |
| url | https://doi.org/10.1038/s41467-024-45987-5 |
| work_keys_str_mv | AT xieshutingdeng ananticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT weisun ananticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT xueyanli ananticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT jiuyuwang ananticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT zhicheng ananticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT gangsheng ananticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT yanliwang ananticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT xieshutingdeng anticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT weisun anticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT xueyanli anticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT jiuyuwang anticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT zhicheng anticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT gangsheng anticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding AT yanliwang anticrisprthatrepressesitsowntranscriptionwhileblockingcas9targetdnabinding |