Site specificity and chromatographic properties of <i>E. COLI</i> K12 and <i>Eco</i>RII DNA‐cytosine methylases

Buryanov, Yа. I.; Bogdarina, Irina; Bayev, A.A.

doi:10.1016/0014-5793(78)80186-0

Cited by 27 publications

(7 citation statements)

References 13 publications

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“…Synthetic double-strand DNA encoding a translational start ATG and viral (SV40) nuclear localization signal (NLS) was then ligated upstream of and in frame with M.EcoRII. The properties of the expressed M.EcoRII were normal as determined by the methods described previously (35). …”

Section: Methodsmentioning

confidence: 99%

Transgene-induced CCWGG methylation does not alter CG methylation patterning in human kidney cells

Шевчук¹

2005

Nucleic Acids Research

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Several reports suggest that CmCWGG methylation tends not to co-exist with mCG methylation in human cells. We have asked whether or not methylation at CCWGG sites can influence CG methylation. DNA from cells expressing an M.EcoRII–GFP fusion was actively methylated at CCWGG sites. CG methylation as measured by R.HpaII/R.MspI ratios was unchanged in cells expressing the transgene. Cloned representatives of CmCWGG methylated DNA often contained, or were adjacent to an ALU repeat, suggesting that M.EcoRII-GFP actively methylated gene-rich R-band DNA. The transgenic methyltransferase applied CmCWGG methylation to a representative human promoter that was heavily methylated at CG dinucleotides (the SERPINB5 promoter) and to a representative promoter that was essentially unmethylated at CG dinucleotides (the APC promoter). In each case, the CG methylation pattern remained in its original state, unchanged by the presence of neighboring CmCWGG sites. Q-PCR measurements showed that RNA expression from the APC gene was not significantly altered by the presence of CmCWGG in its promoter. Kinetic studies suggested that an adjacent CmCWGG methylation site influences neither the maintenance nor the de novo methylation activities of purified human Dnmt1. We conclude that CmCWGG methylation does not exert a significant effect on CG methylation in human kidney cells.

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Section: Methodsmentioning

confidence: 99%

Transgene-induced CCWGG methylation does not alter CG methylation patterning in human kidney cells

Шевчук¹

2005

Nucleic Acids Research

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“…EcoRII and BstNI DNA MTases can be also used for probing CpNpG-motif methylation in the CCWGG sequences of eukaryotic DNA [39]. Both enzymes recognize this duplex sequence and methylate inner cytosine residue in it to generate different products [40,41]. EcoRII DNA MTase methylates the cytosine residues at position C5 of the pyrimidine base to give C5-methylcytosine, whereas BstNI DNA MTase methylates these residues at the exocyclic amino group to give N 4 -methylcytosine.…”

Section: Ccwgg Methylationmentioning

confidence: 99%

The use of prokaryotic DNA methyltransferases as experimental and analytical tools in modern biology

Buryanov

Шевчук

2005

Analytical Biochemistry

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“…To explore the function of the selfish element and to test if it has undergone pseudogenization, we cloned the Kat-HpaII-CM and its closest bacterial homologue in terms of sequence identity, Algibacter HpaII-CM (Alg-HpaII-CM), into plasmid pACYC184 and expressed them in E. coli Top10 cells. This E. coli strain does not contain any methyltransferases that target the putative HpaII-CM recognition sequence (CCGG), but instead expresses Dcm methylase, which methylates the second cytosine residue in CCWGG [28], and Dam methylase, which methylates adenine residues in the sequence GATC [29]. The HpaII-CM-expressing E. coli Top10 strains were cultured for 16 h alongside a control strain harbouring an empty plasmid.…”

Section: Resultsmentioning

confidence: 99%

A functional bacterial-derived restriction modification system in the mitochondrion of a heterotrophic protist

Milner

Wideman

Stairs

et al. 2021

Preprint

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The overarching trend in mitochondrial evolution is functional streamlining coupled with gene loss; therefore, gene acquisition by mitochondria is considered to be exceedingly rare. Selfish elements in the form of self-splicing introns occur in many organellar genomes, but the wider diversity of selfish elements, and how they persist in organellar genomes, has not been explored. In the mitochondrial genome of a marine heterotrophic katablepharid protist, we identify a functional type II restriction modification system originating from a horizontal gene transfer event involving bacteria related to flavobacteria. This restriction modification system consists of an HpaII-like endonuclease and a cognate cytosine methyltransferase. We demonstrate that these proteins are functional by heterologous expression in both bacterial and eukaryotic cells. These results suggest that toxin-antitoxin selfish elements, such as restriction modification systems, could be co-opted by eukaryotic genomes to drive uniparental organellar inheritance.

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Site specificity and chromatographic properties of E. COLI K12 and EcoRII DNA‐cytosine methylases

Cited by 27 publications

References 13 publications

Transgene-induced CCWGG methylation does not alter CG methylation patterning in human kidney cells

Transgene-induced CCWGG methylation does not alter CG methylation patterning in human kidney cells

The use of prokaryotic DNA methyltransferases as experimental and analytical tools in modern biology

A functional bacterial-derived restriction modification system in the mitochondrion of a heterotrophic protist

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