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.的II型DNA甲基转移酶的特性分析

Characterisation of Type II DNA Methyltransferases of .

作者信息

Vogelgsang Lars, Nisar Azlan, Scharf Sebastian Alexander, Rommerskirchen Anna, Belick Dana, Dilthey Alexander, Henrich Birgit

机构信息

Institute of Medical Microbiology and Hospital Hygiene, Medical Faculty of the Heinrich-Heine-University Duesseldorf, Universitätsstraße 1, 40225 Duesseldorf, Germany.

出版信息

Microorganisms. 2023 Jun 15;11(6):1591. doi: 10.3390/microorganisms11061591.

DOI:10.3390/microorganisms11061591
PMID:37375093
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10305163/
Abstract

Bacterial virulence, persistence and defence are affected by epigenetic modifications, including DNA methylation. Solitary DNA methyltransferases modulate a variety of cellular processes and influence bacterial virulence; as part of a restriction-modification (RM) system, they act as a primitive immune system in methylating the own DNA, while unmethylated foreign DNA is restricted. We identified a large family of type II DNA methyltransferases in , comprising six solitary methyltransferases and four RM systems. Motif-specific 5mC and 6mA methylations were identified with a tailored Tombo analysis on Nanopore reads. Selected motifs with methylation scores >0.5 fit with the gene presence of DAM1 and DAM2, DCM2, DCM3, and DCM6, but not for DCM1, whose activity was strain-dependent. The activity of DCM1 for CCWGG and of both DAM1 and DAM2 for GATC was proven in methylation-sensitive restriction and finally for recombinant rDCM1 and rDAM2 against a -, -negative background. A hitherto unknown 8/3 gene fusion containing a (TA) repeat region of varying length was characterized within a single strain, suggesting the expression of DCM8/DAM3 phase variants. The combination of genetic, bioinformatics, and enzymatic approaches enabled the detection of a huge family of type II DNA MTases in , whose involvement in virulence and defence can now be characterized in future work.

摘要

细菌的毒力、持久性和防御能力受表观遗传修饰（包括DNA甲基化）的影响。单个DNA甲基转移酶可调节多种细胞过程并影响细菌毒力；作为限制修饰（RM）系统的一部分，它们在甲基化自身DNA时充当原始免疫系统，而未甲基化的外来DNA则受到限制。我们在[具体物种]中鉴定出一个II型DNA甲基转移酶大家族，其中包括六个单个甲基转移酶和四个RM系统。通过对纳米孔读数进行定制的Tombo分析，鉴定出了基序特异性的5mC和6mA甲基化。甲基化得分>0.5的选定基序与DAM1和DAM2、DCM2、DCM3和DCM6的基因存在相符，但与DCM1不符，其活性具有菌株依赖性。通过甲基化敏感限制证明了DCM1对CCWGG的活性以及DAM1和DAM2对GATC的活性，最终在针对[具体菌株]的重组rDCM1和rDAM2、[具体菌株]阴性背景下得到了验证。在单个菌株中鉴定出一个迄今未知的包含不同长度（TA）重复区域的8/3基因融合体，这表明DCM8/DAM3相变体的表达。遗传、生物信息学和酶学方法的结合使得能够在[具体物种]中检测到一个庞大的II型DNA甲基转移酶家族，其在毒力和防御中的作用现在可以在未来的研究中进行表征。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/ed8203f3bea3/microorganisms-11-01591-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/0a3162c921da/microorganisms-11-01591-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/5d53b8db2cd8/microorganisms-11-01591-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/8713156f7580/microorganisms-11-01591-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/7f7a09dc4e26/microorganisms-11-01591-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/312a566a491f/microorganisms-11-01591-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/202d2383614f/microorganisms-11-01591-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/db3715e3c240/microorganisms-11-01591-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/48b964e88c07/microorganisms-11-01591-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/68f624905dcc/microorganisms-11-01591-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/ed8203f3bea3/microorganisms-11-01591-g010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/0a3162c921da/microorganisms-11-01591-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/5d53b8db2cd8/microorganisms-11-01591-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/8713156f7580/microorganisms-11-01591-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/7f7a09dc4e26/microorganisms-11-01591-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/312a566a491f/microorganisms-11-01591-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/202d2383614f/microorganisms-11-01591-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/db3715e3c240/microorganisms-11-01591-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/48b964e88c07/microorganisms-11-01591-g008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/68f624905dcc/microorganisms-11-01591-g009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1844/10305163/ed8203f3bea3/microorganisms-11-01591-g010.jpg

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