Protospacer Adjacent Motif-Induced Allostery Activates CRISPR-Cas9

Palermo, Giulia; Ricci, Clarisse G.; Fernando, Amendra; Basak, Rajshekhar; Jinek, Martin; Rivalta, Ivan; Batista, Victor S.; McCammon, J. Andrew

doi:10.1021/jacs.7b05313

Title: Protospacer Adjacent Motif-Induced Allostery Activates CRISPR-Cas9

Journal Article · 02 August 2017 · Journal of the American Chemical Society

DOI: https://doi.org/10.1021/jacs.7b05313 · OSTI ID:1489021

^[1];

^[2]; Basak, Rajshekhar ^[2]; Jinek, Martin ^[3];

^[4];

^[2]; McCammon, J. Andrew ^[1]

Univ. of California San Diego, La Jolla, CA (United States)
Yale Univ., New Haven, CT (United States)
Univ. of Zurich, Zurich (Switzerland)
Univ. Claude Bernard Lyon, Lyon (France)

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. As a result, targeting such an allosteric mechanism should enable control of CRISPR-Cas9 functionality.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States). National Energy Research Scientific Computing Center (NERSC)

Sponsoring Organization:: USDOE

OSTI ID:: 1489021

Journal Information:: Journal of the American Chemical Society, Vol. 139, Issue 45; ISSN 0002-7863

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 83 works

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