Work

Conformational states of Cas9-sgRNA-DNA ternary complex in the presence of magnesium

Year: 2019

Type: dataset

Abstract: . In the precatalytic state, Cas9 adopts the 'checkpoint' conformation with the HNH nuclease domain positioned far away from the DNA. Transition to the postcatalytic state involves a dramatic ~34-Å swing of the HNH domain and disorder of the REC2 recognition domain. The postcatalytic state captures the cleaved substrate bound to the catalytically competent HNH active site. In the product state, the HNH domain is disordered, REC2 returns to the precatalytic conformation, and additional interactions of REC3 and RuvC with nucleic acids are formed. The coupled domain motions and interactions between the enzyme and the RNA-DNA hybrid provide new insights into the mechanism of genome editing by Cas9.

Source: Worldwide Protein Data Bank

DOI: 10.2210/pdb6o0z/pdb

Authors: X. Zhu, R. Clarke, A.K. Puppala, S. Chittori, A. Merk, B.J. Merrill, M. Simonovic, S. Subramaniam

Institutions: National Institutes of Health, National Cancer Institute, Center for Cancer Research, University of Illinois Chicago, University of British Columbia

Language: English

Topic: CRISPR and Genetic Engineering

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: National Institutes of Health, Eunice Kennedy Shriver National Institute of Child Health and Human Development

Awards: Technology Development for 3D Electron Microscopy, Studies on the mechanisms governing selenium incorporation into proteins, Wnt/Beta-catenin and Tcf/Lef factor regulation of pluripotency

OpenAlex ID: W2959956949

Cites: 32

Cited by: 81

Related to: 20