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Title: A Functional Mini-Integrase in a Two-Protein Type V-C CRISPR System

Abstract

CRISPR-Cas immunity requires integration of short, foreign DNA fragments into the host genome at the CRISPR locus, a site consisting of alternating repeat sequences and foreign-derived spacers. In most CRISPR systems, the proteins Cas1 and Cas2 form the integration complex and are both essential for DNA acquisition. Most type V-C and V-D systems lack the cas2 gene and have unusually short CRISPR repeats and spacers. Here, we show that a mini-integrase comprising the type V-C Cas1 protein alone catalyzes DNA integration with a preference for short (17- to 19-base-pair) DNA fragments. The mini-integrase has weak specificity for the CRISPR array. We present evidence that the Cas1 proteins form a tetramer for integration. Our findings support a model of a minimal integrase with an internal ruler mechanism that favors shorter repeats and spacers. This minimal integrase may represent the function of the ancestral Cas1 prior to Cas2 adoption.

Authors:
 [1];  [2];  [3];  [2];  [4];  [4];  [5];  [6];  [7]
  1. Univ. of California, Berkeley, CA (United States); Harvard Univ., Cambridge, MA (United States)
  2. Univ. of California, Berkeley, CA (United States)
  3. Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences (QB3); Tel Aviv Univ., Ramat Aviv (Israel)
  4. USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
  5. Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences (QB3)
  6. Univ. of California, Berkeley, CA (United States). Innovative Genomics Inst.
  7. Univ. of California, Berkeley, CA (United States). Innovative Genomics Inst., Howard Hughes Medical Inst.; Gladstone Inst., San Francisco, CA (United States)
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC); National Science Foundation (NSF); National Institutes of Health (NIH)
OSTI Identifier:
1581329
Grant/Contract Number:  
AC02-05CH11231; 1S10OD020062–01; 1244557; T32 066698
Resource Type:
Accepted Manuscript
Journal Name:
Molecular Cell
Additional Journal Information:
Journal Volume: 73; Journal Issue: 4; Journal ID: ISSN 1097-2765
Publisher:
Cell Press - Elsevier
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Wright, Addison V., Wang, Joy Y., Burstein, David, Harrington, Lucas B., Paez-Espino, David, Kyrpides, Nikos C., Iavarone, Anthony T., Banfield, Jillian F., and Doudna, Jennifer A. A Functional Mini-Integrase in a Two-Protein Type V-C CRISPR System. United States: N. p., 2019. Web. doi:10.1016/j.molcel.2018.12.015.
Wright, Addison V., Wang, Joy Y., Burstein, David, Harrington, Lucas B., Paez-Espino, David, Kyrpides, Nikos C., Iavarone, Anthony T., Banfield, Jillian F., & Doudna, Jennifer A. A Functional Mini-Integrase in a Two-Protein Type V-C CRISPR System. United States. doi:10.1016/j.molcel.2018.12.015.
Wright, Addison V., Wang, Joy Y., Burstein, David, Harrington, Lucas B., Paez-Espino, David, Kyrpides, Nikos C., Iavarone, Anthony T., Banfield, Jillian F., and Doudna, Jennifer A. Tue . "A Functional Mini-Integrase in a Two-Protein Type V-C CRISPR System". United States. doi:10.1016/j.molcel.2018.12.015. https://www.osti.gov/servlets/purl/1581329.
@article{osti_1581329,
title = {A Functional Mini-Integrase in a Two-Protein Type V-C CRISPR System},
author = {Wright, Addison V. and Wang, Joy Y. and Burstein, David and Harrington, Lucas B. and Paez-Espino, David and Kyrpides, Nikos C. and Iavarone, Anthony T. and Banfield, Jillian F. and Doudna, Jennifer A.},
abstractNote = {CRISPR-Cas immunity requires integration of short, foreign DNA fragments into the host genome at the CRISPR locus, a site consisting of alternating repeat sequences and foreign-derived spacers. In most CRISPR systems, the proteins Cas1 and Cas2 form the integration complex and are both essential for DNA acquisition. Most type V-C and V-D systems lack the cas2 gene and have unusually short CRISPR repeats and spacers. Here, we show that a mini-integrase comprising the type V-C Cas1 protein alone catalyzes DNA integration with a preference for short (17- to 19-base-pair) DNA fragments. The mini-integrase has weak specificity for the CRISPR array. We present evidence that the Cas1 proteins form a tetramer for integration. Our findings support a model of a minimal integrase with an internal ruler mechanism that favors shorter repeats and spacers. This minimal integrase may represent the function of the ancestral Cas1 prior to Cas2 adoption.},
doi = {10.1016/j.molcel.2018.12.015},
journal = {Molecular Cell},
number = 4,
volume = 73,
place = {United States},
year = {2019},
month = {1}
}

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Works referencing / citing this record:

Adaptation processes that build CRISPR immunity: creative destruction, updated
journal, June 2019

  • Lau, Chun H.; Reeves, Ryan; Bolt, Edward L.
  • Essays in Biochemistry, Vol. 63, Issue 2
  • DOI: 10.1042/ebc20180073

Sequence motifs recognized by the casposon integrase of Aciduliprofundum boonei
journal, May 2019

  • Béguin, Pierre; Chekli, Yankel; Sezonov, Guennadi
  • Nucleic Acids Research, Vol. 47, Issue 12
  • DOI: 10.1093/nar/gkz447