Molecular characterization of the DNA methyltransferase M1.NcuI from Neisseria cuniculi ATCC 14688

Furmanek, B; Sektas, Marian; Wons, Ewa; Kaczorowski, Tadeusz

doi:10.1016/j.resmic.2006.10.006

Cited by 6 publications

(12 citation statements)

References 53 publications

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“…In the case of M.MspI the enzyme dimerizes at higher protein concentrations, which may reflect a tendency to aggregate (Dubey et al, 1992). This is in agreement with our earlier finding that methyltransferases M1.MboII and M1.NcuI purified from overexpressing clones exist in both monomeric and dimeric forms (Furmanek et al, 2007).…”

Section: Enzymic Properties Of M2mboiisupporting

confidence: 92%

“…Protein concentrations were determined by the method of Bradford (1976), with BSA as the standard. Overexpression and purification of M1.MboII and M1.NcuI have been described previously (Furmanek et al, 2007).…”

Section: Methodsmentioning

confidence: 99%

“…Our studies have also been motivated by the fact that the MboII R-M system from M. bovis (Brown et al, 1980) has the same sequence specificity as another studied in our laboratory, the NcuI R-M system from Neisseria cuniculi ATCC 14688 (Furmanek et al, 2007). It was of especial interest to find out what isospecific R-M enzymes have in common at the structural and functional level.…”

Section: Introductionmentioning

confidence: 94%

“…Isospecific systems MboII and NcuI have a similar genetic organization (Furmanek et al, 2007). However, in contrast to the rather compact gene structure of the NcuI R-M system, the two MboII methyltransferases are separated by an intergenic region of 684 bp (Fig.…”

Section: Protein Sequence Analysis and Specificity Of Mboii Methyltramentioning

confidence: 99%

“…It has been shown recently that the substrate specificity and the level of activity for two separately existing methyltransferases may be strikingly different, even though they modify either the same or different nucleotides of target DNA (Cerritelli et al, 1989;Merkiene et al, 1998;Kriukiene et al, 2005). The MboII R-M system as well as its isospecific systems NcuI (Furmanek et al, 2007) and HpyAII (Lin et al, 2001), and other homologous R-M systems from Helicobacter pylori strains (Ando et al, 2000;Xu et al, 2000), are members of the IIS class, which generally require two methyltransferase activities, one for each strand of their asymmetrical recognition sites. In this case, the nucleotide sequence of the recognition site has profound significance.…”

Section: Introductionmentioning

confidence: 99%

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M1.MboII and M2.MboII type IIS methyltransferases: different specificities, the same target

et al. 2009

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Methylation of a base in a specific DNA sequence protects the DNA from nucleolytic cleavage by restriction enzymes recognizing the same sequence. The MboII restriction-modification (R-M) system of Moraxella bovis ATCC 10900 consists of a restriction endonuclease gene and two methyltransferase genes. The enzymes encoded by this system recognize an asymmetrical sequence 59-GAAGA-39/39-CTTCT-59. M1.MboII modifies the last adenine in the recognition sequence 59-GAAGA-39 to N 6 -methyladenine. A second methylase, M2.MboII, was cloned and purified to electrophoretic homogeneity using a four-step chromatographic procedure. It was demonstrated that M2.MboII modifies the internal cytosine in the recognition sequence 39-CTTCT-59, yielding N 4 -methylcytosine, and moreover is able to methylate single-stranded DNA. ) inhibit M2.MboII methylation activity. It was found that the isomethylomer M2.NcuI from Neisseria cuniculi ATCC 14688 behaves in the same manner. Functional analysis showed that the complete MboII R-M system, consisting of two methyltransferases genes and the mboIIR gene, is the most stable and the least harmful to bacterial cells.

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Section: Enzymic Properties Of M2mboiisupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 94%

Section: Protein Sequence Analysis and Specificity Of Mboii Methyltramentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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M1.MboII and M2.MboII type IIS methyltransferases: different specificities, the same target

et al. 2009

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Evaluation of the enzymatic properties of DNA (cytosine-5)-methyltransferase M.ApeKI from archaea in the presence of metal ions

Hayashi,

Wada,

Yamamura

et al. 2024

Bioscience, Biotechnology, and Biochemistry

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We previously identified M.ApeKI from Aeropyum pernix K1 as a highly thermostable DNA (cytosine-5)-methyltransferase. M.ApeKI uses the type II restriction-modification system (R-M system), among the best-studied R-M systems. Although endonucleases generally utilize Mg (II) as a cofactor, several reports have shown that MTases exhibit different reactions in the presence of metal ions. This study aim was to evaluate the enzymatic properties of DNA (cytosine-5)-methyltransferase M.ApeKI from archaea in the presence of metal ions. We evaluated the influence of metal ions on the catalytic activity and DNA binding of M.ApeKI. The catalytic activity was inhibited by Cu (II), Mg (II), Mn (II), and Zn (II), each at 5 mM. DNA binding was more strongly inhibited by 5 mM Cu (II) and 10 mM Zn (II). To our knowledge, this is the first report showing that DNA binding of typeII MTase is inhibited by metal ions.

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RNA editing by T7 RNA polymerase bypasses InDel mutations causing unexpected phenotypic changes

Wons

Furmanek-Blaszk

Sektas

2015

Nucleic Acids Research

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DNA-dependent T7 RNA polymerase (T7 RNAP) is the most powerful tool for both gene expression and in vitro transcription. By using a Next Generation Sequencing (NGS) approach we have analyzed the polymorphism of a T7 RNAP-generated mRNA pool of the mboIIM2 gene. We find that the enzyme displays a relatively high level of template-dependent transcriptional infidelity. The nucleotide misincorporations and multiple insertions in A/T-rich tracts of homopolymers in mRNA (0.20 and 0.089%, respectively) cause epigenetic effects with significant impact on gene expression that is disproportionally high to their frequency of appearance. The sequence-dependent rescue of single and even double InDel frameshifting mutants and wild-type phenotype recovery is observed as a result. As a consequence, a heterogeneous pool of functional and non-functional proteins of almost the same molecular mass is produced where the proteins are indistinguishable from each other upon ordinary analysis. We suggest that transcriptional infidelity as a general feature of the most effective RNAPs may serve to repair and/or modify a protein function, thus increasing the repertoire of phenotypic variants, which in turn has a high evolutionary potential.

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Molecular characterization of the DNA methyltransferase M1.NcuI from Neisseria cuniculi ATCC 14688

Cited by 6 publications

References 53 publications

M1.MboII and M2.MboII type IIS methyltransferases: different specificities, the same target

M1.MboII and M2.MboII type IIS methyltransferases: different specificities, the same target

Evaluation of the enzymatic properties of DNA (cytosine-5)-methyltransferase M.ApeKI from archaea in the presence of metal ions

RNA editing by T7 RNA polymerase bypasses InDel mutations causing unexpected phenotypic changes

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