[23] Characterization of type II DNA-methyltransferases

Landry, David; Barsomian, Janet M.; Feehery, George R.; Wilson, Geoffrey G.

doi:10.1016/0076-6879(92)16025-f

Cited by 11 publications

(5 citation statements)

References 42 publications

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“…Samples (5 mg) of the synthetic double-stranded ODN were methylated as described for MTase assays and separated on a denaturing polyacrylamide gel (Landry et al, 1992). The bands were excised, and the gel slices were crushed in microfuge tubes and incubated overnight at room temperature in 300 ml 0?5 M ammonium acetate, 1 mM EDTA.…”

Section: Methodsmentioning

confidence: 99%

DNA methyltransferases from Neisseria meningitidis and Neisseria gonorrhoeae FA1090 associated with mismatch nicking endonucleases

et al. 2004

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The genes encoding the DNA methyltransferases M.NmeDI and M.NmeAI from Neisseria meningitidis associated with the genes encoding putative Vsr endonucleases were overexpressed in Escherichia coli. The enzymes were purified to apparent homogeneity on Ni-NTA agarose columns, yielding proteins of 49+/-1 kDa and 39.6+/-1 kDa, respectively, under denaturing conditions. M.NmeDI recognizes the degenerate sequence 5'-RCCGGB-3'. It methylates the first 5' cytosine residue on both strands within the core sequence CCGG. The enzyme shows higher affinity with the hemimethylated degenerate sequence than with the unmethylated degenerate sequence. Comparison of the amino acid sequence of the target-recognizing domain of M.NmeDI with the closest neighbours recognizing the sequence 5'-RCCGGY-3' showed the presence of the homologous domain and an additional domain that may be responsible for recognizing the degenerate sequence. M.NmeAI recognizes the sequence 5'-CCGG-3' and methylates the second 5' cytosine residue on both DNA strands. In Neisseria gonorrhoeae strain FA1090 the homologues of these ORFs are truncated due to a variety of mutations.

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Section: Methodsmentioning

confidence: 99%

DNA methyltransferases from Neisseria meningitidis and Neisseria gonorrhoeae FA1090 associated with mismatch nicking endonucleases

et al. 2004

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“…Although the methylated components can be detected and quantified using a variety of chromatographic techniques (2,(7)(8)(9), their localization in longer stretches of DNA has long been a formidable task (1, 10 -12; reviewed in 13). The development of a genomic sequencing technique that affords a "positive" display of m5C residues on dideoxy-sequencing ladders has been an important advance in the field of DNA analysis (14).…”

Section: Laboratory Of Biological Dna Modification Institute Of Biotmentioning

confidence: 99%

Bisulfite Sequencing Protocol Displays both 5-Methylcytosine and N4-Methylcytosine

Vilkaitis¹,

Klimašauskas²

1999

Analytical Biochemistry

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“…DNA R-M systems have been classified as Type I, II, IIs, and III based on cofactor requirements, protein structure, and the nature of the recognition cut sites. Type II systems are found in most bacteria and are the simplest in structure, containing a methyltransferase (MTase) and a restriction endonuclease (ENase) that act independently of one another (3,4). Type IIs systems differ from Type II systems in that their recognition sequences are non-palindromic and the scission event does not take place within the recognition sequence (5).…”

Section: Introductionmentioning

confidence: 99%

The Neisseria gonorrhoeae S.NgoVIII restriction/modification system: a type IIs system homologous to the Haemophilus parahaemolyticus HphI restriction/modification system

Stein

1997

Nucleic Acids Research

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Strains of Neisseria gonorrhoeae possess numerous restriction-modification (R-M) systems. One of these systems, which has been found in all strains tested, encodes the S. NgoVIII specificity (5'TCACC 3') R-M system. We cloned two adjacent methyltransferase genes (dcmH and damH), each encoding proteins whose actions protect DNA from digestion by R.HphI or R.Ngo BI (5'TCACC 3'). The damH gene product is a N 6-methyladenine methyltransferase that recognizes this sequence. We constructed a plasmid containing multiple copies of the S.NgoVIII sequence, grew it in the presence of damH and used the HPLC to demonstrate the presence of N 6-methyladenine in the DNA. A second plasmid, containing overlapping damH and Escherichia coli dam recognition sequences in combination with various restriction digests, was used to identify which adenine in the recognition sequence was modified by damH. The predicted dcmH gene product is homologous to 5-methylcytosine methyltransferases. The products of both the dcmH and damH genes, as well as an open reading frame downstream of the damH gene are highly similar to the Haemophilus parahaemolyticus hphIMC , hphIMA and hphIR gene products, encoding the Hph I Type IIs R-M system. The S.NgoVIII R-M genes are flanked by a 97 bp direct repeat that may be involved in the mobility of this R-M system.

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[23] Characterization of type II DNA-methyltransferases

Cited by 11 publications

References 42 publications

DNA methyltransferases from Neisseria meningitidis and Neisseria gonorrhoeae FA1090 associated with mismatch nicking endonucleases

DNA methyltransferases from Neisseria meningitidis and Neisseria gonorrhoeae FA1090 associated with mismatch nicking endonucleases

Bisulfite Sequencing Protocol Displays both 5-Methylcytosine and N4-Methylcytosine

The Neisseria gonorrhoeae S.NgoVIII restriction/modification system: a type IIs system homologous to the Haemophilus parahaemolyticus HphI restriction/modification system

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