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A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro

Deazaguanine DNA modifications are widespread in phages, particularly in those with pathogenic hosts. Pseudomonas phage iggy substitutes ∼16.5% of its genomic 2′-deoxyguanosine (G) with dPreQ(0), and the iggy deazaguanine transglycosylase (DpdA) is unique in having a strict GA target motif, not obse...

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Autores principales: Olsen, Nikoline S, Nielsen, Tue K, Cui, Liang, Dedon, Peter, Neve, Horst, Hansen, Lars H, Kot, Witold
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10484667/
https://www.ncbi.nlm.nih.gov/pubmed/37503841
http://dx.doi.org/10.1093/nar/gkad622
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author Olsen, Nikoline S
Nielsen, Tue K
Cui, Liang
Dedon, Peter
Neve, Horst
Hansen, Lars H
Kot, Witold
author_facet Olsen, Nikoline S
Nielsen, Tue K
Cui, Liang
Dedon, Peter
Neve, Horst
Hansen, Lars H
Kot, Witold
author_sort Olsen, Nikoline S
collection PubMed
description Deazaguanine DNA modifications are widespread in phages, particularly in those with pathogenic hosts. Pseudomonas phage iggy substitutes ∼16.5% of its genomic 2′-deoxyguanosine (G) with dPreQ(0), and the iggy deazaguanine transglycosylase (DpdA) is unique in having a strict GA target motif, not observed previously. The iggy PreQ(0) modification is shown to provide protection against both restriction endonucleases and Cas9 (when present in PAM), thus expanding our understanding of the deazaguanine modification system, its potential, and diversity. Phage iggy represents a new genus of Pseudomonas phages within the Queuovirinae subfamily; which have very little in common with other published phage genomes in terms of nucleotide similarity (<10%) and common proteins (<2%). Interestingly, shared similarity is concentrated in dpdA and preQ(0) biosynthesis genes. TEM imaging confirmed a siphovirus morphology with a prolate icosahedral head and a non-contractile flexible tail with one long central tail spike. The observed protective effect of the deazaguanine modification on the iggy DNA may contribute to its broad within-species host range. Phage iggy was isolated on Pseudomonas aeruginosa PAO1, but also infects PDO300, PAK, PA14, as well as 10 of 27 tested environmental isolates and 13 of 20 tested clinical isolates of P. aeruginosa from patients with cystic fibrosis.
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spelling pubmed-104846672023-09-08 A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro Olsen, Nikoline S Nielsen, Tue K Cui, Liang Dedon, Peter Neve, Horst Hansen, Lars H Kot, Witold Nucleic Acids Res Molecular Biology Deazaguanine DNA modifications are widespread in phages, particularly in those with pathogenic hosts. Pseudomonas phage iggy substitutes ∼16.5% of its genomic 2′-deoxyguanosine (G) with dPreQ(0), and the iggy deazaguanine transglycosylase (DpdA) is unique in having a strict GA target motif, not observed previously. The iggy PreQ(0) modification is shown to provide protection against both restriction endonucleases and Cas9 (when present in PAM), thus expanding our understanding of the deazaguanine modification system, its potential, and diversity. Phage iggy represents a new genus of Pseudomonas phages within the Queuovirinae subfamily; which have very little in common with other published phage genomes in terms of nucleotide similarity (<10%) and common proteins (<2%). Interestingly, shared similarity is concentrated in dpdA and preQ(0) biosynthesis genes. TEM imaging confirmed a siphovirus morphology with a prolate icosahedral head and a non-contractile flexible tail with one long central tail spike. The observed protective effect of the deazaguanine modification on the iggy DNA may contribute to its broad within-species host range. Phage iggy was isolated on Pseudomonas aeruginosa PAO1, but also infects PDO300, PAK, PA14, as well as 10 of 27 tested environmental isolates and 13 of 20 tested clinical isolates of P. aeruginosa from patients with cystic fibrosis. Oxford University Press 2023-07-28 /pmc/articles/PMC10484667/ /pubmed/37503841 http://dx.doi.org/10.1093/nar/gkad622 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Molecular Biology
Olsen, Nikoline S
Nielsen, Tue K
Cui, Liang
Dedon, Peter
Neve, Horst
Hansen, Lars H
Kot, Witold
A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro
title A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro
title_full A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro
title_fullStr A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro
title_full_unstemmed A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro
title_short A novel Queuovirinae lineage of Pseudomonas aeruginosa phages encode dPreQ(0) DNA modifications with a single GA motif that provide restriction and CRISPR Cas9 protection in vitro
title_sort novel queuovirinae lineage of pseudomonas aeruginosa phages encode dpreq(0) dna modifications with a single ga motif that provide restriction and crispr cas9 protection in vitro
topic Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10484667/
https://www.ncbi.nlm.nih.gov/pubmed/37503841
http://dx.doi.org/10.1093/nar/gkad622
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