The DOI matches its Crossref record; 5 of the 5 references it cites were found in a registry or an index.
Structures of CRISPR Cas3 offer mechanistic insights into Cascade-activated DNA unwinding and degradation
Open accessAbstract
CRISPR drives prokaryotic adaptation to invasive nucleic acids such as phages and plasmids, using an RNA-mediated interference mechanism. Interference in type I CRISPR-Cas systems requires a targeting Cascade complex and a degradation machine, Cas3, which contains both nuclease and helicase activities. Here we report the crystal structures of Thermobifida fusca Cas3 bound to single-stranded (ss) DNA substrate and show that it is an obligate 3′-to-5′ ssDNase that preferentially accepts substrate directly from the helicase moiety. Conserved residues in the HD-type nuclease coordinate two irons for ssDNA cleavage. We demonstrate ATP coordination and conformational flexibility of the SF2-type helicase domain. Cas3 is specifically guided toward Cascade-bound target DNA by a PAM sequence, through physical interactions with both the nontarget substrate strand and the CasA protein. The sequence of recognition events ensures well-controlled DNA targeting and degradation of foreign DNA by Cascade and Cas3.
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An updated evolutionary classification of CRISPR–Cas systems
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Biology and Applications of CRISPR Systems: Harnessing Nature’s Toolbox for Genome Engineering
Wright, A. V., Nuñez, J. K., & Doudna, J. A. (2016) · Cell · doi:10.1016/j.cell.2015.12.035 · Cited by 989
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Metadata
- Authors
- Huo, Y., Nam, K. H., Ding, F., Lee, H., Wu, L., Xiao, Y., Farchione, M. D., Zhou, S., Rajashankar, K., Kurinov, I., Zhang, R., & Ke, A.
- Venue
- Nature Structural & Molecular Biology
- Published
- 2014-08-17
- Volume
- 21
- Issue
- 9
- Pages
- 771-777
- ISSN
- 1545-9985 (ISSN-L)
- 1545-9993
- OpenAlex
- W1996555720
- Cited by
- 186
- References
- 5
- FWCI
- 6.81
- Type
- journal article
- Language
- en
- Open access
- green
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- Huo, Y., Nam, K. H., Ding, F., Lee, H., Wu, L., Xiao, Y., Farchione, M. D., Zhou, S., Rajashankar, K., Kurinov, I., Zhang, R., & Ke, A. (2014). Structures of CRISPR Cas3 offer mechanistic insights into Cascade-activated DNA unwinding and degradation. Nature Structural & Molecular Biology. https://doi.org/10.1038/nsmb.2875
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