Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA

Abstract E. coli phage 9 g contains the modified base deoxyarchaeosine (dG+) in its genome. The phage encodes its own primase, DNA ligase, DNA polymerase, and enzymes necessary to synthesize and incorporate dG+. Here we report phage 9 g DNA sensitivity to >200 Type II restriction endonucleases (R...

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Autores principales:	Rebecca Tsai, Ivan R. Corrêa, Michael Y. Xu, Shuang-yong Xu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/96957818cd1641e88ff5a1e64be62e66
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spelling	oai:doaj.org-article:96957818cd1641e88ff5a1e64be62e662021-12-02T15:05:27ZRestriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA10.1038/s41598-017-08864-42045-2322https://doaj.org/article/96957818cd1641e88ff5a1e64be62e662017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08864-4https://doaj.org/toc/2045-2322Abstract E. coli phage 9 g contains the modified base deoxyarchaeosine (dG+) in its genome. The phage encodes its own primase, DNA ligase, DNA polymerase, and enzymes necessary to synthesize and incorporate dG+. Here we report phage 9 g DNA sensitivity to >200 Type II restriction endonucleases (REases). Among the REases tested approximately 29% generated complete or partial digestions, while the remaining 71% displayed resistance to restriction. Phage 9 g restriction fragments can be degraded by DNA exonucleases or ligated by T3 and T4 DNA ligases. In addition, we examined a number of cytosine and adenine methyltransferases to generate double base modifications. M.AluI, M.CviPI, M.HhaI, and M.EcoGII were able to introduce 5mC or N6mA into 9 g DNA as confirmed by partial resistance to restriction and by liquid chromatography-mass spectrometry. A number of wild-type E. coli bacteria restricted phage 9 g, indicating natural restriction barriers exist in some strains. A BlastP search of GenBank sequences revealed five glutamine amidotransferase-QueC homologs in Enterobacteria and Pseudomonas phage, and distant homologs in other phage and bacterial genomes, suggesting that dG+ is not a rare modification. We also mapped phage 9 g DNA packaging (pac) site containing two 21-bp direct repeats and a major terminase cleavage site in the phage genome.Rebecca TsaiIvan R. CorrêaMichael Y. XuShuang-yong XuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-13 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rebecca Tsai Ivan R. Corrêa Michael Y. Xu Shuang-yong Xu Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA
description	Abstract E. coli phage 9 g contains the modified base deoxyarchaeosine (dG+) in its genome. The phage encodes its own primase, DNA ligase, DNA polymerase, and enzymes necessary to synthesize and incorporate dG+. Here we report phage 9 g DNA sensitivity to >200 Type II restriction endonucleases (REases). Among the REases tested approximately 29% generated complete or partial digestions, while the remaining 71% displayed resistance to restriction. Phage 9 g restriction fragments can be degraded by DNA exonucleases or ligated by T3 and T4 DNA ligases. In addition, we examined a number of cytosine and adenine methyltransferases to generate double base modifications. M.AluI, M.CviPI, M.HhaI, and M.EcoGII were able to introduce 5mC or N6mA into 9 g DNA as confirmed by partial resistance to restriction and by liquid chromatography-mass spectrometry. A number of wild-type E. coli bacteria restricted phage 9 g, indicating natural restriction barriers exist in some strains. A BlastP search of GenBank sequences revealed five glutamine amidotransferase-QueC homologs in Enterobacteria and Pseudomonas phage, and distant homologs in other phage and bacterial genomes, suggesting that dG+ is not a rare modification. We also mapped phage 9 g DNA packaging (pac) site containing two 21-bp direct repeats and a major terminase cleavage site in the phage genome.
format	article
author	Rebecca Tsai Ivan R. Corrêa Michael Y. Xu Shuang-yong Xu
author_facet	Rebecca Tsai Ivan R. Corrêa Michael Y. Xu Shuang-yong Xu
author_sort	Rebecca Tsai
title	Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA
title_short	Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA
title_full	Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA
title_fullStr	Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA
title_full_unstemmed	Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA
title_sort	restriction and modification of deoxyarchaeosine (dg+)-containing phage 9 g dna
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/96957818cd1641e88ff5a1e64be62e66
work_keys_str_mv	AT rebeccatsai restrictionandmodificationofdeoxyarchaeosinedgcontainingphage9gdna AT ivanrcorrea restrictionandmodificationofdeoxyarchaeosinedgcontainingphage9gdna AT michaelyxu restrictionandmodificationofdeoxyarchaeosinedgcontainingphage9gdna AT shuangyongxu restrictionandmodificationofdeoxyarchaeosinedgcontainingphage9gdna
_version_	1718388847513960448

Restriction and modification of deoxyarchaeosine (dG+)-containing phage 9 g DNA

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