Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble
Abstract
Type III-A CRISPR-Cas systems are prokaryotic RNA-guided adaptive immune systems that use a protein-RNA complex, Csm, for transcription-dependent immunity against foreign DNA. Csm can cleave RNA and single-stranded DNA (ssDNA), but whether it targets one or both nucleic acids during transcription elongation is unknown. Here, we show that binding of a Thermus thermophilus (T. thermophilus) Csm (TthCsm) to a nascent transcript in a transcription elongation complex (TEC) promotes tethering but not direct contact of TthCsm with RNA polymerase (RNAP). Biochemical experiments show that both TthCsm and Staphylococcus epidermidis (S. epidermidis) Csm (SepCsm) cleave RNA transcripts, but not ssDNA, at the transcription bubble. Taken together, these results suggest that Type III systems primarily target transcripts, instead of unwound ssDNA in TECs, for immunity against double-stranded DNA (dsDNA) phages and plasmids. This reveals similarities between Csm and eukaryotic RNA interference, which also uses RNA-guided RNA targeting to silence actively transcribed genes.
- Authors:
-
[1];
[2];
[2]
- Univ. of California, Berkeley, CA (United States)
- Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC)
- OSTI Identifier:
- 1559231
- Grant/Contract Number:
- AC02-05CH11231
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Nature Communications
- Additional Journal Information:
- Journal Volume: 10; 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
Liu, Tina Y., Liu, Jun-Jie, Aditham, Abhishek J., Nogales, Eva, and Doudna, Jennifer A.. Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble. United States: N. p., 2019.
Web. doi:10.1038/s41467-019-10780-2.
Liu, Tina Y., Liu, Jun-Jie, Aditham, Abhishek J., Nogales, Eva, & Doudna, Jennifer A.. Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble. United States. https://doi.org/10.1038/s41467-019-10780-2
Liu, Tina Y., Liu, Jun-Jie, Aditham, Abhishek J., Nogales, Eva, and Doudna, Jennifer A.. Fri .
"Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble". United States. https://doi.org/10.1038/s41467-019-10780-2. https://www.osti.gov/servlets/purl/1559231.
@article{osti_1559231,
title = {Target preference of Type III-A CRISPR-Cas complexes at the transcription bubble},
author = {Liu, Tina Y. and Liu, Jun-Jie and Aditham, Abhishek J. and Nogales, Eva and Doudna, Jennifer A.},
abstractNote = {Type III-A CRISPR-Cas systems are prokaryotic RNA-guided adaptive immune systems that use a protein-RNA complex, Csm, for transcription-dependent immunity against foreign DNA. Csm can cleave RNA and single-stranded DNA (ssDNA), but whether it targets one or both nucleic acids during transcription elongation is unknown. Here, we show that binding of a Thermus thermophilus (T. thermophilus) Csm (TthCsm) to a nascent transcript in a transcription elongation complex (TEC) promotes tethering but not direct contact of TthCsm with RNA polymerase (RNAP). Biochemical experiments show that both TthCsm and Staphylococcus epidermidis (S. epidermidis) Csm (SepCsm) cleave RNA transcripts, but not ssDNA, at the transcription bubble. Taken together, these results suggest that Type III systems primarily target transcripts, instead of unwound ssDNA in TECs, for immunity against double-stranded DNA (dsDNA) phages and plasmids. This reveals similarities between Csm and eukaryotic RNA interference, which also uses RNA-guided RNA targeting to silence actively transcribed genes.},
doi = {10.1038/s41467-019-10780-2},
journal = {Nature Communications},
number = 1,
volume = 10,
place = {United States},
year = {2019},
month = {7}
}
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