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Structural insights into the apo-structure of Cpf1 protein from Francisella novicida.
- Source :
-
Biochemical and biophysical research communications [Biochem Biophys Res Commun] 2018 Apr 15; Vol. 498 (4), pp. 775-781. Date of Electronic Publication: 2018 Mar 12. - Publication Year :
- 2018
-
Abstract
- Clustered regularly interspaced short palindromic repeats (CRISPRs) from Prevotella and Francisella 1 (Cpf1) are RNA-guided endonucleases that produce cohesive double-stranded breaks in DNA by specifically recognizing thymidine-rich protospacer-adjacent motif (PAM) sequences. Cpf1 is emerging as a powerful genome-editing tool. Despite previous structural studies on various Cpf1 proteins, the apo-structure of Cpf1 remains unknown. In the present study, we determined the solution structure of the Cpf1 protein from Francisella novicida (FnCpf1) with and without CRISPR RNA (crRNA) using small-angle X-ray scattering, providing the insights into the apo-structure of FnCpf1. The apo-structure of FnCpf1 was also visualized using negative staining electron microcopy. When we compared the apo-structure of FnCpf1 with crRNA-bound structure, their overall shapes (a closed form) were similar, suggesting that conformational change upon crRNA binding to FnCpf1 is not drastic, but a local induced fit might occur to recognize PAM sequences. In contrast, the apo Cpf1 from Moraxella bovoculi 237 (MbCpf1) was analyzed as an open form, implying that a large conformational change from an open to a closed form might be required for crRNA binding to MbCpf1. These results suggested that the crRNA-induced conformational changes in Cpf1 differ among species.<br /> (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Subjects :
- Bacterial Proteins ultrastructure
CRISPR-Associated Proteins ultrastructure
Clustered Regularly Interspaced Short Palindromic Repeats
Crystallography, X-Ray
Endonucleases ultrastructure
Models, Molecular
Protein Conformation
RNA-Binding Proteins ultrastructure
Bacterial Proteins chemistry
CRISPR-Associated Proteins chemistry
Endonucleases chemistry
Francisella chemistry
RNA-Binding Proteins chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1090-2104
- Volume :
- 498
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Biochemical and biophysical research communications
- Publication Type :
- Academic Journal
- Accession number :
- 29526756
- Full Text :
- https://doi.org/10.1016/j.bbrc.2018.03.057