Structural coordination between active sites of a CRISPR reverse transcriptase-integrase complex
Abstract
CRISPR-Cas systems provide adaptive immunity in bacteria and archaea, beginning with integration of foreign sequences into the host CRISPR genomic locus and followed by transcription and maturation of CRISPR RNAs (crRNAs). In some CRISPR systems, a reverse transcriptase (RT) fusion to the Cas1 integrase and Cas6 maturase creates a single protein that enables concerted sequence integration and crRNA production. To elucidate how the RT-integrase organizes distinct enzymatic activities, we present the cryo-EM structure of a Cas6-RT-Cas1—Cas2 CRISPR integrase complex. The structure reveals a heterohexamer in which the RT directly contacts the integrase and maturase domains, suggesting functional coordination between all three active sites. Together with biochemical experiments, our data support a model of sequential enzymatic activities that enable CRISPR sequence acquisition from RNA and DNA substrates. These findings highlight an expanded capacity of some CRISPR systems to acquire diverse sequences that direct CRISPR-mediated interference.
- Authors:
-
[2];
[4];
[2];
[5]
- Univ. of California, Berkeley, CA (United States). Dept. of Chemistry, California Inst. for Quantitative Biosciences
- Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences, Dept. of Molecular and Cell Biology, Helen Wills Neuroscience Inst.
- Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences, Dept. of Plant and Microbial Biology
- Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology, Dept. of Earth and Planetary Sciences, Dept. of Environmental Science, Policy, and Management
- Univ. of California, Berkeley, CA (United States). Dept. of Chemistry. California Inst. for Quantitative Biosciences. Dept. of Molecular and Cell Biology. Innovative Genomics Inst. Howard Hughes Medical Inst.; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging Division; Gladstone Inst., San Francisco, CA (United States). Gladstone Inst. of Data Science and Biotechnology
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1816040
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Nature Communications
- Additional Journal Information:
- Journal Volume: 12; Journal Issue: 1; Journal ID: ISSN 2041-1723
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Wang, Joy Y., Hoel, Christopher M., Al-Shayeb, Basem, Banfield, Jillian F., Brohawn, Stephen G., and Doudna, Jennifer A. Structural coordination between active sites of a CRISPR reverse transcriptase-integrase complex. United States: N. p., 2021.
Web. doi:10.1038/s41467-021-22900-y.
Wang, Joy Y., Hoel, Christopher M., Al-Shayeb, Basem, Banfield, Jillian F., Brohawn, Stephen G., & Doudna, Jennifer A. Structural coordination between active sites of a CRISPR reverse transcriptase-integrase complex. United States. https://doi.org/10.1038/s41467-021-22900-y
Wang, Joy Y., Hoel, Christopher M., Al-Shayeb, Basem, Banfield, Jillian F., Brohawn, Stephen G., and Doudna, Jennifer A. Thu .
"Structural coordination between active sites of a CRISPR reverse transcriptase-integrase complex". United States. https://doi.org/10.1038/s41467-021-22900-y. https://www.osti.gov/servlets/purl/1816040.
@article{osti_1816040,
title = {Structural coordination between active sites of a CRISPR reverse transcriptase-integrase complex},
author = {Wang, Joy Y. and Hoel, Christopher M. and Al-Shayeb, Basem and Banfield, Jillian F. and Brohawn, Stephen G. and Doudna, Jennifer A.},
abstractNote = {CRISPR-Cas systems provide adaptive immunity in bacteria and archaea, beginning with integration of foreign sequences into the host CRISPR genomic locus and followed by transcription and maturation of CRISPR RNAs (crRNAs). In some CRISPR systems, a reverse transcriptase (RT) fusion to the Cas1 integrase and Cas6 maturase creates a single protein that enables concerted sequence integration and crRNA production. To elucidate how the RT-integrase organizes distinct enzymatic activities, we present the cryo-EM structure of a Cas6-RT-Cas1—Cas2 CRISPR integrase complex. The structure reveals a heterohexamer in which the RT directly contacts the integrase and maturase domains, suggesting functional coordination between all three active sites. Together with biochemical experiments, our data support a model of sequential enzymatic activities that enable CRISPR sequence acquisition from RNA and DNA substrates. These findings highlight an expanded capacity of some CRISPR systems to acquire diverse sequences that direct CRISPR-mediated interference.},
doi = {10.1038/s41467-021-22900-y},
journal = {Nature Communications},
number = 1,
volume = 12,
place = {United States},
year = {Thu May 06 00:00:00 EDT 2021},
month = {Thu May 06 00:00:00 EDT 2021}
}
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