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A reverse transcriptase-Cas1 fusion protein contains a Cas6 domain required for both CRISPR RNA biogenesis and RNA spacer acquisition

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dc.contributor.authorMohr, Georg
dc.contributor.authorSilas, Sukrit
dc.contributor.authorStamos, Jennifer L.
dc.contributor.authorMakarova, Kira S.
dc.contributor.authorMarkham, Laura M.
dc.contributor.authorYao, Jun
dc.contributor.authorElío Lucas, Patrícia
dc.contributor.authorSánchez Amat, Antonio
dc.contributor.authorFire, Andrew Z.
dc.contributor.authorKoonin, Eugene V.
dc.contributor.authorLambowitz, Alan M.
dc.contributor.departmentGenética y Microbiología
dc.date.accessioned2026-01-12T13:07:16Z
dc.date.available2026-01-12T13:07:16Z
dc.date.copyright© 2018 Elsevier Inc.
dc.date.issued2018-11-15
dc.description.abstractProkaryotic CRISPR-Cas systems provide adaptive immunity by integrating portions of foreign nucleic acids (spacers) into genomic CRISPR arrays. Cas6 proteins then process CRISPR array transcripts into spacer-derived RNAs (CRISPR RNAs; crRNAs) that target Cas nucleases to matching invaders. We find that a Marinomonas mediterranea fusion protein combines three enzymatic domains (Cas6, reverse transcriptase [RT], and Cas1), which function in both crRNA biogenesis and spacer acquisition from RNA and DNA. We report a crystal structure of this divergent Cas6, identify amino acids required for Cas6 activity, show that the Cas6 domain is required for RT activity and RNA spacer acquisition, and demonstrate that CRISPR-repeat binding to Cas6 regulates RT activity. Co-evolution of putative interacting surfaces suggests a specific structural interaction between the Cas6 and RT domains, and phylogenetic analysis reveals repeated, stable association of free-standing Cas6s with CRISPR RTs in multiple microbial lineages, indicating that a functional interaction between these proteins preceded evolution of the fusion.
dc.formatapplication/pdf
dc.format.extent24
dc.identifier.citationMolecular Cell, 2018, Vol. 72, Issue 4, pp. 700-714.e8
dc.identifier.doihttps://doi.org/10.1016/j.molcel.2018.09.013
dc.identifier.eissn1097-4164
dc.identifier.issn1097-2765
dc.identifier.urihttp://hdl.handle.net/10201/185889
dc.languageeng
dc.publisherCell Press
dc.relationThis work was supported by NIH R01 grants GM37706 to A.Z.F. and GM37949 to A.M.L. S.S. was supported by a Stanford Graduate Fellowship and an HHMI International Student Research Fellowship. K.S.M. and E.V.K. are supported by the Intramural Program of the U.S. Department of Health and Human Services (via funds provided to the National Library of Medicine). A.S.-A. received funding from the Spanish “Ministerio de Economia, Industria y Competitividad” under project BFU2017-85464 supported by FEDER funds. This research used resources of the Advanced Light Source, which is a DOE Office of Science User Facility under contract no. DE-AC02-05CH11231.
dc.relation.publisherversionhttps://www.sciencedirect.com/science/article/pii/S1097276518307846
dc.rights.accessRightsinfo:eu-repo/semantics/restrictedAccess
dc.subject.odsNo relacionado con ningún objetivo de desarrollo sostenible
dc.titleA reverse transcriptase-Cas1 fusion protein contains a Cas6 domain required for both CRISPR RNA biogenesis and RNA spacer acquisition
dc.typeinfo:eu-repo/semantics/article
dc.type.versioninfo:eu-repo/semantics/publishedVersion
dspace.entity.typePublicationes
relation.isAuthorOfPublication66095fe4-223f-4dbc-9b73-7cade64d2e20
relation.isAuthorOfPublication15331c24-15e1-4518-824f-57238c36ed42
relation.isAuthorOfPublication.latestForDiscovery66095fe4-223f-4dbc-9b73-7cade64d2e20
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