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Detection of preQ0 deazaguanine modifications in bacteriophage CAjan DNA using Nanopore sequencing reveals same hypermodification at two distinct DNA motifs
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gkaa735.pdf
Description
Published version
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6.24 MB
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Adobe PDF
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4216f4696a1256ad61d4988413102124
Author(s) • • • • • • • • •
Kot, Witold
Olsen, Nikoline S
Nielsen, Tue K
Hutinet, Geoffrey
de Crécy-Lagard, Valérie
Cui, Liang
Dedon, Peter C
Carstens, Alexander B
Moineau, Sylvain
Swairjo, Manal A
Date Issued
2020
Journal
Nucleic Acids Research
Publisher
Oxford University Press (OUP)
Version
Final published version
Abstract
© 2020 The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. In the constant evolutionary battle against mobile genetic elements (MGEs), bacteria have developed several defense mechanisms, some of which target the incoming, foreign nucleic acids e.g. restriction-modification (R-M) or CRISPR-Cas systems. Some of these MGEs, including bacteriophages, have in turn evolved different strategies to evade these hurdles. It was recently shown that the siphophage CAjan and 180 other viruses use 7-deazaguanine modifications in their DNA to evade bacterial R-M systems. Among others, phage CAjan genome contains a gene coding for a DNA-modifying homolog of a tRNA-deazapurine modification enzyme, together with four 7-cyano-7-deazaguanine synthesis genes. Using the CRISPR-Cas9 genome editing tool combined with the Nanopore Sequencing (ONT) we showed that the 7-deazaguanine modification in the CAjan genome is dependent on phage-encoded genes. The modification is also site-specific and is found mainly in two separate DNA sequence contexts: GA and GGC. Homology modeling of the modifying enzyme DpdA provides insight into its probable DNA binding surface and general mode of DNA recognition.
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Creative Commons Attribution NonCommercial License 4.0
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DOI of Published Version
10.1093/NAR/GKAA735