{"publisher":"Cold Spring Harbor Laboratory","date_published":"2022-06-14T00:00:00Z","year":"2022","month":"06","language":[{"iso":"eng"}],"title":"Assembly of multi-subunit fusion proteins into the RNA-targeting type III-D CRISPR-Cas effector complex","_id":"17116","type":"preprint","oa_version":"Preprint","article_processing_charge":"No","publication_status":"submitted","user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","status":"public","day":"14","date_updated":"2024-06-04T06:58:41Z","main_file_link":[{"url":"https://doi.org/10.1101/2022.06.13.496011","open_access":"1"}],"citation":{"ama":"Schwartz EA, Bravo JPK, Macias LA, et al. Assembly of multi-subunit fusion proteins into the RNA-targeting type III-D CRISPR-Cas effector complex. <i>bioRxiv</i>. doi:<a href=\"https://doi.org/10.1101/2022.06.13.496011\">10.1101/2022.06.13.496011</a>","mla":"Schwartz, Evan A., et al. “Assembly of Multi-Subunit Fusion Proteins into the RNA-Targeting Type III-D CRISPR-Cas Effector Complex.” <i>BioRxiv</i>, Cold Spring Harbor Laboratory, doi:<a href=\"https://doi.org/10.1101/2022.06.13.496011\">10.1101/2022.06.13.496011</a>.","apa":"Schwartz, E. A., Bravo, J. P. K., Macias, L. A., McCafferty, C. L., Dangerfield, T. L., Walker, J. N., … Taylor, D. W. (n.d.). Assembly of multi-subunit fusion proteins into the RNA-targeting type III-D CRISPR-Cas effector complex. <i>bioRxiv</i>. Cold Spring Harbor Laboratory. <a href=\"https://doi.org/10.1101/2022.06.13.496011\">https://doi.org/10.1101/2022.06.13.496011</a>","ista":"Schwartz EA, Bravo JPK, Macias LA, McCafferty CL, Dangerfield TL, Walker JN, Brodbelt JS, Fineran PC, Fagerlund RD, Taylor DW. Assembly of multi-subunit fusion proteins into the RNA-targeting type III-D CRISPR-Cas effector complex. bioRxiv, <a href=\"https://doi.org/10.1101/2022.06.13.496011\">10.1101/2022.06.13.496011</a>.","ieee":"E. A. Schwartz <i>et al.</i>, “Assembly of multi-subunit fusion proteins into the RNA-targeting type III-D CRISPR-Cas effector complex,” <i>bioRxiv</i>. Cold Spring Harbor Laboratory.","chicago":"Schwartz, Evan A., Jack Peter Kelly Bravo, Luis A. Macias, Caitlyn L. McCafferty, Tyler L. Dangerfield, Jada N. Walker, Jennifer S. Brodbelt, Peter C. Fineran, Robert D. Fagerlund, and David W. Taylor. “Assembly of Multi-Subunit Fusion Proteins into the RNA-Targeting Type III-D CRISPR-Cas Effector Complex.” <i>BioRxiv</i>. Cold Spring Harbor Laboratory, n.d. <a href=\"https://doi.org/10.1101/2022.06.13.496011\">https://doi.org/10.1101/2022.06.13.496011</a>.","short":"E.A. Schwartz, J.P.K. Bravo, L.A. Macias, C.L. McCafferty, T.L. Dangerfield, J.N. Walker, J.S. Brodbelt, P.C. Fineran, R.D. Fagerlund, D.W. Taylor, BioRxiv (n.d.)."},"publication":"bioRxiv","extern":"1","abstract":[{"text":"CRISPR (Clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems are a type of adaptive immune response in bacteria and archaea that utilize crRNA (CRISPR RNA)-guided effector complexes to target complementary RNA or DNA for destruction. The prototypical type III-A and III-B CRISPR-Cas systems utilize multi-subunit effector complexes composed of individual proteins to cleave ssRNA targets at 6-nt intervals, as well as non-specifically degrading ssDNA and activating cyclic oligoadenylate (cOA) synthesis. Recent studies have shown that type III systems can contain subunit fusions yet maintain canonical type III RNA-targeting capabilities. To understand how a multi-subunit fusion effector functions, we determine structures of a variant type III-D effector and biochemically characterize how it cleaves RNA targets. These findings provide insights into how multi-subunit fusion proteins are tethered together and assemble into an active and programmable RNA endonuclease, how the effector utilizes a novel mechanism for target RNA seeding, and the structural basis for the evolution of type III effector complexes. Furthermore, our results provide a blueprint for fusing subunits in class 1 effectors for design of user-defined effector complexes with disparate activities.</jats:p><jats:sec><jats:title>Important note</jats:title><jats:p>While this manuscript was in preparation, a manuscript describing the structure of the type III-E effector was published<jats:sup>1</jats:sup>. We reference these important findings; however, a careful comparison of the structures will follow once the coordinates have been released by the PDB.","lang":"eng"}],"author":[{"full_name":"Schwartz, Evan A.","first_name":"Evan A.","last_name":"Schwartz"},{"orcid":"0000-0003-0456-0753","last_name":"Bravo","full_name":"Bravo, Jack Peter Kelly","first_name":"Jack Peter Kelly","id":"96aecfa5-8931-11ee-af30-aa6a5d6eee0e"},{"last_name":"Macias","first_name":"Luis A.","full_name":"Macias, Luis A."},{"first_name":"Caitlyn L.","full_name":"McCafferty, Caitlyn L.","last_name":"McCafferty"},{"last_name":"Dangerfield","full_name":"Dangerfield, Tyler L.","first_name":"Tyler L."},{"last_name":"Walker","first_name":"Jada N.","full_name":"Walker, Jada N."},{"full_name":"Brodbelt, Jennifer S.","first_name":"Jennifer S.","last_name":"Brodbelt"},{"full_name":"Fineran, Peter C.","first_name":"Peter C.","last_name":"Fineran"},{"first_name":"Robert D.","full_name":"Fagerlund, Robert D.","last_name":"Fagerlund"},{"last_name":"Taylor","first_name":"David W.","full_name":"Taylor, David W."}],"oa":1,"date_created":"2024-06-04T06:44:16Z","doi":"10.1101/2022.06.13.496011"}