Dataset Information

Enhanced Cleavage at Abasic Sites within Clustered Lesions in Nucleosome Core Particles.

ABSTRACT: Clustered lesions are a hallmark of ?-radiolysis, but are produced by other damaging agents as well. Bistranded clustered lesions are precursors to double-strand breaks and are challenging to repair, thus making them an especially deleterious form of DNA damage. An abasic site (AP) is an alkaline-labile lesion frequently present in clustered lesions. Strand scission at an AP site is accelerated ?100-fold in nucleosome core particles (NCPs). We examined how AP reactivity was affected within clustered lesions in NCPs. The rate constant of strand scission is increased as much as 2.5-fold in the presence of a proximal abasic site or thymidine glycol in the complementary strand. A proximal mispair has a similar effect on AP reactivity. Increased AP reactivity within a clustered lesion correlates with decreased UV melting temperatures of the corresponding duplexes compared to one containing an isolated abasic site. However, the thermodynamics of duplex melting do not correlate with AP reactivity within different clustered lesions. Overall, increased AP reactivity within clustered lesions is attributed to greater access of histone proteins to the lesion due to decreased duplex stability.

SUBMITTER: Yang K

PROVIDER: S-EPMC6172155 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature

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Publications

Enhanced Cleavage at Abasic Sites within Clustered Lesions in Nucleosome Core Particles.

Yang Kun K Greenberg Marc M MM

Chembiochem : a European journal of chemical biology 20180824 19

Clustered lesions are a hallmark of γ-radiolysis, but are produced by other damaging agents as well. Bistranded clustered lesions are precursors to double-strand breaks and are challenging to repair, thus making them an especially deleterious form of DNA damage. An abasic site (AP) is an alkaline-labile lesion frequently present in clustered lesions. Strand scission at an AP site is accelerated ≈100-fold in nucleosome core particles (NCPs). We examined how AP reactivity was affected within clust ...[more]

PMID: 30043401

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Project description:DNA is damaged by various endogenous and exogenous sources, leading to a diverse group of reactive intermediates that yield a complex mixture of products. The initially formed products are often metastable and can react to yield lesions that are more biologically deleterious. Mechanistic studies are frequently carried out on free DNA as the substrate. The observations do not necessarily reflect the reaction environment inside human cells where genomic DNA is condensed as chromatin in the nucleus. Chromatin is made up of monomeric structural units called nucleosomes, which are comprised of DNA wrapped around an octameric core of histone proteins (two copies each of histones H2A, H2B, H3, and H4).This account presents a summary of our work in the past decade on the mechanistic studies of DNA damage and repair in reconstituted nucleosome core particles (NCPs). A series of metastable lesions and reactive intermediates, such as abasic sites (AP), N7-methyl-2'-deoxyguanosine (MdG), and 2'-deoxyadenosin-N6-yl radical (dA•), have been independently generated in a site-specific manner in bottom-up-synthesized NCPs. Detailed mechanistic studies on these NCPs revealed that histones actively participate in DNA damage and repair processes in diverse ways. For instance, nucleophilic residues in the flexible histone N-terminal tails, such as Lys and N-terminal α-amine, react with electrophilic DNA damage and reactive intermediates. In some cases, transient intermediates are produced, leading to the promotion or suppression of damage and repair processes. In other examples, reactions with histones yield reversible or stable DNA-protein cross-links (DPCs). Histones also utilize acidic and basic residues, such as histidine and aspartic acid, to catalyze DNA strand cleavage through general acid/base catalysis. Alternatively, a Tyr in histone plays a vital role in nucleosomal DNA damage and repair via radical transfer. Finally, the reactivity discovered during the mechanistic studies has facilitated the development of new reagents and methods with applications in biotechnology.This research has enriched our knowledge of the roles of histone proteins in DNA damage and repair and their contributions to epigenetics and may have significant biological implications. The residues in histone N-terminal tails that react with DNA lesions also play pivotal roles in regulating the structure and function of chromatin, indicating that there may be cross-talk between DNA damage and repair in eukaryotic cells and epigenetic regulation. Also, in view of the biased amino acid composition of histones, these results provide hints about how the proteins have evolved to minimize their deleterious effects but maximize beneficial ones for maintaining genome integrity. Finally, previously unreported DPCs and histone post-translational modifications have been discovered through this research. The effects of these newly identified lesions on the structure and function of chromatin and their fates inside cells remain to be elucidated.

Dataset Information

Enhanced Cleavage at Abasic Sites within Clustered Lesions in Nucleosome Core Particles.

Publications

Enhanced Cleavage at Abasic Sites within Clustered Lesions in Nucleosome Core Particles.

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