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

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.

doi:10.1016/j.molcel.2018.12.015

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

Journal Article · Tue Jan 29 00:00:00 EST 2019 · Molecular Cell

DOI:https://doi.org/10.1016/j.molcel.2018.12.015· OSTI ID:1581329

Wright, Addison V. ^[1]; Wang, Joy Y. ^[2]; Burstein, David ^[3]; Harrington, Lucas B. ^[2]; Paez-Espino, David ^[4]; Kyrpides, Nikos C. ^[4]; Iavarone, Anthony T. ^[5]; Banfield, Jillian F. ^[6]; Doudna, Jennifer A. ^[7]

Univ. of California, Berkeley, CA (United States); Harvard Univ., Cambridge, MA (United States)
Univ. of California, Berkeley, CA (United States)
Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences (QB3); Tel Aviv Univ., Ramat Aviv (Israel)
USDOE Joint Genome Institute (JGI), Walnut Creek, CA (United States)
Univ. of California, Berkeley, CA (United States). California Inst. for Quantitative Biosciences (QB3)
Univ. of California, Berkeley, CA (United States). Innovative Genomics Inst.
Univ. of California, Berkeley, CA (United States). Innovative Genomics Inst., Howard Hughes Medical Inst.; Gladstone Inst., San Francisco, CA (United States)

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.

View Accepted Manuscript (DOE)

Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); National Science Foundation (NSF); USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1581329

Journal Information:: Molecular Cell, Journal Name: Molecular Cell Journal Issue: 4 Vol. 73; ISSN 1097-2765

Publisher:: Cell Press - ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Sequence motifs recognized by the casposon integrase of Aciduliprofundum boonei Béguin, Pierre; Chekli, Yankel; Sezonov, Guennadi Nucleic Acids Research, Vol. 47, Issue 12 https://doi.org/10.1093/nar/gkz447	journal	May 2019
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Casposase structure and the mechanistic link between DNA transposition and spacer acquisition by CRISPR-Cas Hickman, Alison B.; Kailasan, Shweta; Genzor, Pavol eLife, Vol. 9 https://doi.org/10.7554/elife.50004	journal	January 2020

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