Dataset Information

Slow Rotational Dynamics of Cytosine NH₂ Groups in Double-Stranded DNA.

ABSTRACT: Aromatic NH₂ groups are essential as hydrogen-bond donors in secondary structures of DNA and RNA. Although rapid rotations of NH₂ groups of adenine and guanine bases were previously characterized, there has been a lack of quantitative information about slow rotations of cytosine NH₂ groups in Watson-Crick base pairs. In this study, using an NMR method we had recently developed, we determined the kinetic rate constants and energy barriers for cytosine NH₂ rotations in a 15-base-pair DNA duplex. Our data show that the rotational dynamics of cytosine NH₂ groups depend on local environments. Qualitative correlation between the ranges of ¹⁵N chemical shifts and rotational time scales for various NH₂ groups of nucleic acids and proteins illuminates a relationship between the partial double-bond character of the C-N bond and the time scale for NH₂ rotations.

SUBMITTER: Wang X

PROVIDER: S-EPMC9805297 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Slow Rotational Dynamics of Cytosine NH2 Groups in Double-Stranded DNA.

Wang Xi X Yu Binhan B Iwahara Junji J

Biochemistry 20220627 14

Aromatic NH2 groups are essential as hydrogen-bond donors in secondary structures of DNA and RNA. Although rapid rotations of NH2 groups of adenine and guanine bases were previously characterized, there has been a lack of quantitative information about slow rotations of cytosine NH2 groups in Watson-Crick base pairs. In this study, using an NMR method we had recently developed, we determined the kinetic rate constants and energy barriers for cytosine NH2</su ...[more]

PMID: 35759792

Similar Datasets

Project description:Methylation of cytosine bases is strongly linked to gene expression, imprinting, aging, and carcinogenesis. The Ten-eleven translocation (TET) family of enzymes, which are Fe(II)/2-oxoglutarate (2OG)-dependent enzymes, employ Fe(IV)=O species to dealkylate the lesioned bases to an unmodified cytosine. Recently, it has been shown that the TET2 enzyme can catalyze promiscuously DNA substrates containing unnatural alkylated cytosine. Such unnatural substrates of TET can be used as direct probes for measuring the TET activity or capturing TET from cellular samples. Herein, we studied the catalytic mechanisms during the oxidation of the unnatural C5-position modifications (5-ethylcytosine (5eC), 5-vinylcytosine (5vC) and 5-ethynylcytosine (5eyC)) and the demethylation of N4-methylated lesions (4-methylcytosine (4mC) and 4,4-dimethylcytosine(4dmC)) of the cytosine base by the TET2 enzyme using molecular dynamics (MD) and combined quantum mechanics and molecular mechanics (QM/MM) computational approaches. The results reveal that the chemical nature of the alkylation of the double-stranded (ds) DNA substrates induces distinct changes in the interactions in the binding site, the second coordination sphere, and long-range correlated motions of the ES complexes. The rate-determining hydrogen atom transfer (HAT) is faster in N4-methyl substituent substrates than in the C5-alkylations. Importantly, the calculations show the preference of hydroxylation over desaturation in both 5eC and 5vC substrates. The studies elucidate the post-hydroxylation rearrangements of the hydroxylated intermediates of 5eyC and 5vC to ketene and 5-formylmethylcytosine (5fmC), respectively, and hydrolysis of hemiaminal intermediate of 4mC to formaldehyde and unmodified cytosine proceed exclusively in aqueous solution outside of the enzyme environment. Overall, the studies show that the chemical nature of the unnatural alkylated cytosine substrates exercises distinct effects on the binding interactions, reaction mechanism, and dynamics of TET2.

Dataset Information

Slow Rotational Dynamics of Cytosine NH₂ Groups in Double-Stranded DNA.

Publications

Slow Rotational Dynamics of Cytosine NH<sub>2</sub> Groups in Double-Stranded DNA.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets