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dc.contributor.authorvan Erp, Paul B. G.
dc.contributor.authorJackson, Ryan N.
dc.contributor.authorCarter, Joshua
dc.contributor.authorGolden, Sarah M.
dc.contributor.authorBailey, Scott
dc.contributor.authorWiedenheft, Blake A.
dc.identifier.citationVan Erp, Paul B.G., Ryan N. Jackson, Joshua Carter, Sarah M. Golden, Scott Bailey, and Blake Wiedenheft. “ Mechanism of CRISPR-RNA Guided Recognition of DNA Targets in Escherichia Coli .” Nucleic Acids Research 43, no. 17 (August 3, 2015): 8381–8391. doi:10.1093/nar/gkv793.en_US
dc.description.abstractIn bacteria and archaea, short fragments of foreign DNA are integrated into Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) loci, providing a molecular memory of previous encounters with foreign genetic elements. In Escherichia coli, short CRISPR-derived RNAs are incorporated into a multi-subunit surveillance complex called Cascade (CRISPR-associated complex for antiviral defense). Recent structures of Cascade capture snapshots of this seahorse-shaped RNA-guided surveillance complex before and after binding to a DNA target. Here we determine a 3.2 Å x-ray crystal structure of Cascade in a new crystal form that provides insight into the mechanism of double-stranded DNA binding. Molecular dynamic simulations performed using available structures reveal functional roles for residues in the tail, backbone and belly subunits of Cascade that are critical for binding double-stranded DNA. Structural comparisons are used to make functional predictions and these predictions are tested in vivo and in vitro. Collectively, the results in this study reveal underlying mechanisms involved in target-induced conformational changes and highlight residues important in DNA binding and protospacer adjacent motif recognition.en_US
dc.description.sponsorshipMSU’s Graduate School and the Office of the Vice President for Research and Economic Development (to P.B.G.v.E.); Ruth L. Kirschstein National Research Service Award from the National Institutes of Health [F32 GM108436 to R.N.J.]; Undergraduate research from the Howard Hughes Medical Institute [#52006931 to J.C.], Montana IDeA Network of Biomedical Research Excellence from the National Institutes of Health [P20GM103474 to J.C.], Irving L. Weissman Undergraduate research (to J.C.); National Institutes of Health [R01GM097330 to S.B.]; National Institutes of Health [P20GM103500, R01GM108888 to B.W.], National Science Foundation EPSCoR [EPS-110134 to B.W.], M.J. Murdock Charitable Trust (to B.W.); Montana State University Agricultural Experimental Station; Advance Photon Source [ACB-12002, AGM-12006, DE-AC02-06CH11357]; Stanford Synchrotron Radiation Lightsource [DE-AC02-76SF00515, P41GM103393]en_US
dc.rightspermits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly citeden_US
dc.titleMechanism of CRISPR-RNA guided recognition of DNA targets in Escherichia colien_US
mus.citation.journaltitleNucleic Acids Researchen_US
mus.identifier.categoryChemical & Material Sciencesen_US
mus.identifier.categoryHealth & Medical Sciencesen_US
mus.relation.collegeCollege of Agricultureen_US
mus.relation.collegeCollege of Letters & Scienceen_US
mus.relation.departmentMicrobiology & Immunology.en_US
mus.relation.universityMontana State University - Bozemanen_US
mus.contributor.orcidCarter, Joshua|0000-0002-9194-7966en_US
mus.contributor.orcidJackson, Ryan N.|0000-0001-7403-3794en_US

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