In vitro Type II Restriction of Bacteriophage DNA With Modified Pyrimidines.
In vitro Type II Restriction of Bacteriophage DNA With Modified Pyrimidines.
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DOI：
10.3389/fmicb.2020.604618
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发表时间：
2020
影响因子：
5.2
通讯作者：
Xu SY
中科院分区：
文献类型：
--
作者：
Flodman K;Corrêa IR Jr;Dai N;Weigele P;Xu SY
为了对抗宿主编码的限制系统，噬菌体（噬菌体）在其基因组中加入修饰的碱基。例如，噬菌体在其基因组中携带修饰的嘧啶，如缺乏α-和β-糖基转移酶的T4gt中的5-羟甲基胞嘧啶（5hmC）， T4中的葡萄糖化5-羟甲基胞嘧啶（5gmC）， Xp12中的5-甲基胞嘧啶（5mC）和SP8中的5-羟甲基脱氧尿嘧啶（5hmdU）。在这项工作中，我们对噬菌体Xp12和SP8基因组进行了测序，并检测了T4gt， T4， Xp12和SP8噬菌体dna的II型限制性。T4gt、T4和Xp12基因组对市售II型限制性内切酶（REases）的抗性分别为81.9%（227种酶中186种）、94.3%（227种酶中214种）和89.9%（218种酶中196种）。然而，SP8基因组对这些酶的抗性仅为~ 8.3%（在所测试的204种酶中有17种）。腺嘌呤DNA甲基转移酶（mtase）如M.Dam和M.EcoGII以及胞嘧啶DNA mtase如CpG甲基化酶可以进一步修饰SP8 DNA。通过5hmdU DNA激酶处理，SP8 DNA中的5hmdU碱基被磷酸化，达到约20%的5hmdU磷酸化，从而对更多的II型限制产生抗性或部分抗性。该工作为分子生物学家处理修饰嘧啶和利用酶提供了方便的参考。噬菌体Xp12和SP8的基因组序列为进一步研究5mC和5hmdU DNA碱基修饰的遗传途径以及比较噬菌体基因组学奠定了基础。
To counteract host-encoded restriction systems, bacteriophages (phages) incorporate modified bases in their genomes. For example, phages carry in their genomes modified pyrimidines such as 5-hydroxymethyl-cytosine (5hmC) in T4gt deficient in α- and β-glycosyltransferases, glucosylated-5-hydroxymethylcytosine (5gmC) in T4, 5-methylcytosine (5mC) in Xp12, and 5-hydroxymethyldeoxyuridine (5hmdU) in SP8. In this work we sequenced phage Xp12 and SP8 genomes and examined Type II restriction of T4gt, T4, Xp12, and SP8 phage DNAs. T4gt, T4, and Xp12 genomes showed resistance to 81.9% (186 out of 227 enzymes tested), 94.3% (214 out of 227 enzymes tested), and 89.9% (196 out of 218 enzymes tested), respectively, commercially available Type II restriction endonucleases (REases). The SP8 genome, however, was resistant to only ∼8.3% of these enzymes (17 out of 204 enzymes tested). SP8 DNA could be further modified by adenine DNA methyltransferases (MTases) such as M.Dam and M.EcoGII as well as a number of cytosine DNA MTases, such as CpG methylase. The 5hmdU base in SP8 DNA was phosphorylated by treatment with a 5hmdU DNA kinase to achieve ∼20% phosphorylated 5hmdU, resulting resistance or partially resistant to more Type II restriction. This work provides a convenient reference for molecular biologists working with modified pyrimidines and using REases. The genomic sequences of phage Xp12 and SP8 lay the foundation for further studies on genetic pathways for 5mC and 5hmdU DNA base modifications and for comparative phage genomics.
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影响因子：
3.2
作者：
Bohacova, Sona;Ludvikova, Lucie;Hocek, Michal
通讯作者：
Hocek, Michal
影响因子：
14.9
作者：
Drozdz, Marek;Piekarowicz, Andrzej;Radlinska, Monika
通讯作者：
Radlinska, Monika
DOI：
10.1073/pnas.1714812115
发表时间：
2018-04-03
影响因子：
11.1
作者：
Lee, Yan-Jiun;Dai, Nan;Weigele, Peter R.
通讯作者：
Weigele, Peter R.
影响因子：
4.6
作者：
He X;Hull V;Thomas JA;Fu X;Gidwani S;Gupta YK;Black LW;Xu SY
通讯作者：
Xu SY
DOI：
10.1073/pnas.62.3.892
发表时间：
1969-01-01
影响因子：
11.1
作者：
GOLD, LM;SCHWEIGER, M
通讯作者：
SCHWEIGER, M