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Structures of the CRISPR genome integration complex.

Wright AV, Liu JJ, Knott GJ, Doxzen KW, Nogales E, Doudna JA
Science 357 1113-1118 (2017)
Related entries: 5vvj, 5vvl, 5wfe

Cited: 78 times
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Abstract

CRISPR-Cas systems depend on the Cas1-Cas2 integrase to capture and integrate short foreign DNA fragments into the CRISPR locus, enabling adaptation to new viruses. We present crystal structures of Cas1-Cas2 bound to both donor and target DNA in intermediate and product integration complexes, as well as a cryo-electron microscopy structure of the full CRISPR locus integration complex, including the accessory protein IHF (integration host factor). The structures show unexpectedly that indirect sequence recognition dictates integration site selection by favoring deformation of the repeat and the flanking sequences. IHF binding bends the DNA sharply, bringing an upstream recognition motif into contact with Cas1 to increase both the specificity and efficiency of integration. These results explain how the Cas1-Cas2 CRISPR integrase recognizes a sequence-dependent DNA structure to ensure site-selective CRISPR array expansion during the initial step of bacterial adaptive immunity.

Articles - 5vvk mentioned but not cited (3)

  1. Structures of the CRISPR genome integration complex. Wright AV, Liu JJ, Knott GJ, Doxzen KW, Nogales E, Doudna JA. Science 357 1113-1118 (2017)
  2. Antibody structure prediction using interpretable deep learning. Ruffolo JA, Sulam J, Gray JJ. Patterns (N Y) 3 100406 (2022)
  3. Casposase structure and the mechanistic link between DNA transposition and spacer acquisition by CRISPR-Cas. Hickman AB, Kailasan S, Genzor P, Haase AD, Dyda F. Elife 9 e50004 (2020)


Reviews citing this publication (18)

  1. Molecular mechanisms of CRISPR-Cas spacer acquisition. McGinn J, Marraffini LA. Nat Rev Microbiol 17 7-12 (2019)
  2. DNA supercoiling and transcription in bacteria: a two-way street. Dorman CJ. BMC Mol Cell Biol 20 26 (2019)
  3. DNA interference and beyond: structure and functions of prokaryotic Argonaute proteins. Lisitskaya L, Aravin AA, Kulbachinskiy A. Nat Commun 9 5165 (2018)
  4. When is a transcription factor a NAP? Dorman CJ, Schumacher MA, Bush MJ, Brennan RG, Buttner MJ. Curr Opin Microbiol 55 26-33 (2020)
  5. Assessment and Comparison of Molecular Subtyping and Characterization Methods for Salmonella. Tang S, Orsi RH, Luo H, Ge C, Zhang G, Baker RC, Stevenson A, Wiedmann M. Front Microbiol 10 1591 (2019)
  6. Structural insights into the inactivation of CRISPR-Cas systems by diverse anti-CRISPR proteins. Zhu Y, Zhang F, Huang Z. BMC Biol 16 32 (2018)
  7. Pruning and Tending Immune Memories: Spacer Dynamics in the CRISPR Array. Garrett SC. Front Microbiol 12 664299 (2021)
  8. Structural biology of CRISPR-Cas immunity and genome editing enzymes. Wang JY, Pausch P, Doudna JA. Nat Rev Microbiol 20 641-656 (2022)
  9. CRISPR-Cas9 in genome editing: Its function and medical applications. Khadempar S, Familghadakchi S, Motlagh RA, Farahani N, Dashtiahangar M, Rezaei H, Gheibi Hayat SM. J Cell Physiol 234 5751-5761 (2019)
  10. Mechanisms of Type I-E and I-F CRISPR-Cas Systems in Enterobacteriaceae. Xue C, Sashital DG. EcoSal Plus 8 (2019)
  11. Molecular-based identification and detection of Salmonella in food production systems: current perspectives. Ricke SC, Kim SA, Shi Z, Park SH. J Appl Microbiol 125 313-327 (2018)
  12. Creating memories: molecular mechanisms of CRISPR adaptation. Lee H, Sashital DG. Trends Biochem Sci 47 464-476 (2022)
  13. Detection of CRISPR adaptation. Shiriaeva A, Fedorov I, Vyhovskyi D, Severinov K. Biochem Soc Trans 48 257-269 (2020)
  14. Adaptation by Type III CRISPR-Cas Systems: Breakthrough Findings and Open Questions. Zhang X, An X. Front Microbiol 13 876174 (2022)
  15. CRISPR-Cas adaptation in Escherichia coli. Mitić D, Bolt EL, Ivančić-Baće I. Biosci Rep 43 BSR20221198 (2023)
  16. CRISPR-Cas-Based Engineering of Probiotics. Liu L, Helal SE, Peng N. Biodes Res 5 0017 (2023)
  17. Molecular Details of DNA Integration by CRISPR-Associated Proteins During Adaptation in Bacteria and Archaea. Flusche T, Rajan R. Adv Exp Med Biol 1414 27-43 (2023)
  18. gRNA Design: How Its Evolution Impacted on CRISPR/Cas9 Systems Refinement. Motoche-Monar C, Ordoñez JE, Chang O, Gonzales-Zubiate FA. Biomolecules 13 1698 (2023)

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