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Protospacer Adjacent Motif-Induced Allostery Activates CRISPR-Cas9.

  • Palermo G Department of Chemistry, Yale University , P.O. Box 208107, New Haven, Connecticut 06520-8107, United States.
  • Ricci CG Department of Chemistry, Yale University , P.O. Box 208107, New Haven, Connecticut 06520-8107, United States.
  • Fernando A Department of Biochemistry, University of Zürich , Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.
  • Basak R Univ Lyon, Ens de Lyon, CNRS, Université Claude Bernard Lyon 1 , Laboratoire de Chimie UMR 5182, F-69342 Lyon, France.
  • Jinek M Department of Chemistry, Yale University , P.O. Box 208107, New Haven, Connecticut 06520-8107, United States.
  • Rivalta I
  • Batista VS
  • McCammon JA
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  • 2017-08-03
Published in:
  • Journal of the American Chemical Society. - 2017
Allosteric Regulation
CRISPR-Associated Proteins
CRISPR-Cas Systems
Catalytic Domain
Clustered Regularly Interspaced Short Palindromic Repeats
DNA Cleavage
Endonucleases
Enzyme Activation
Gene Editing
Molecular Dynamics Simulation
English CRISPR-Cas9 is a genome editing technology with major impact in life sciences. In this system, the endonuclease Cas9 generates double strand breaks in DNA upon RNA-guided recognition of a complementary DNA sequence, which strictly requires the presence of a protospacer adjacent motif (PAM) next to the target site. Although PAM recognition is essential for cleavage, it is unknown whether and how PAM binding activates Cas9 for DNA cleavage at spatially distant sites. Here, we find evidence of a PAM-induced allosteric mechanism revealed by microsecond molecular dynamics simulations. PAM acts as an allosteric effector and triggers the interdependent conformational dynamics of the Cas9 catalytic domains (HNH and RuvC), responsible for concerted cleavage of the two DNA strands. Targeting such an allosteric mechanism should enable control of CRISPR-Cas9 functionality.
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  • English
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hybrid
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https://sonar.ch/global/documents/82333
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