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Mechanism and cleavage specificity of the H-N-H endonuclease colicin E9.
TLDR
The data show that the H-N-H motif is an adaptable catalytic centre able to hydrolyse nucleic acid by different mechanisms depending on the substrate and metal ion regime. Expand
Low-fidelity Pyrococcus furiosus DNA polymerase mutants useful in error-prone PCR.
TLDR
Mutants of the family B DNA polymerase from Pyrococcus furiosus (Pfu-Pol), with superb performance in error-prone PCR, are described, which are anticipated to be extremely useful for the randomization of gene, and hence protein, sequences. Expand
Zebularine: a novel DNA methylation inhibitor that forms a covalent complex with DNA methyltransferases.
TLDR
The interaction between the C5 MTase from Haemophilus haemolyticus and an oligodeoxynucleotide duplex containing 2-H pyrimidinone provides a molecular explanation for the mechanism of action of the anti-cancer drug zebularine. Expand
A read-ahead function in archaeal DNA polymerases detects promutagenic template-strand uracil.
TLDR
DNA polymerases from several hyperthermophilic archaea specifically recognize the presence of uracil in a template strand and stall DNA synthesis before mutagenic misincorporation of adenine, which may represent the first step in a pathway for the repair of cytosine deamination in archaea. Expand
Mechanism-based inhibition of C5-cytosine DNA methyltransferases by 2-H pyrimidinone.
TLDR
It is shown that 2-H pyrimidinone, when incorporated into DNA duplexes containing the recognition sites for M.HgaI-2 and M.MspI, elicits the formation of inhibitory covalent nucleoprotein complexes. Expand
Uracil recognition in archaeal DNA polymerases captured by X-ray crystallography.
TLDR
The crystal structure of V93Q, a polymerase variant that no longer recognises uracil, is reported, explaining the V 93Q phenotype by the steric exclusion of uracic exclusion from the pocket. Expand
Predicting metals sensed by ArsR‐SmtB repressors: allosteric interference by a non‐effector metal
TLDR
Although aseR did not sense zinc in vivo, it bound zinc in vitro exploiting α3 thiols, but AseR DNA binding was not impaired by zinc, so tight non‐effector metal complexes could theoretically inhibit metal sensing. Expand
Structural basis for uracil recognition by archaeal family B DNA polymerases
TLDR
This work has identified a 'pocket' in the N-terminal domains of archaeal DNA polymerases that is positioned to interact with the template strand and provides interacting groups that discriminate uracil from the four normal DNA bases (including thymine). Expand
Site-directed mutagenesis of the catalytic residues of bovine pancreatic deoxyribonuclease I.
TLDR
Results demonstrate that H134, D168, D212 and H252 play a critical role in the catalytic mechanism of DNase I, which uses a two metal-ion hydrolytic mechanism and both magnesium ions are used to supply electrophilic catalysis. Expand
Binding and recognition of GATATC target sequences by the EcoRV restriction endonuclease: a study using fluorescent oligonucleotides and fluorescence polarization.
TLDR
Oligonucleotides labeled with hexachlorofluorescein (hex) have enabled the interaction of the restriction endonuclease EcoRV with DNA to be evaluated using fluorescence anisotropy, and it was found that the ecoRV restriction end onuclease showed a preference of between 1.5 and 6.5 for its GATATC target sequence at pH 7.5. Expand
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