The Base Pairing Partner Modulates Alkylguanine Alkyltransferase

McKeague, Maureen; Otto, Claudia; Räz, Michael H.; Angelov, T.; Sturla, Shana J.

doi:10.1021/acschembio.8b00446

Cited by 4 publications

(3 citation statements)

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“…143 Several biochemical studies have revealed that sequence context, including the opposing base, impacts the repair of O 6 -MedG by AGT. 137,[144][145][146] In contrast, recent work from Essigmann and coworkers showed no specific mutational patterns arising from AGT repair in mouse cells treated with the alkylation agent N-methyl-N-nitrosourea (MNU). 147 However, there is little work on AGT accessibility to different chromatin states in mammalian cells.…”

Section: Methylating Agents (O 6 -Methylguanine)mentioning

confidence: 96%

Next-generation DNA damage sequencing

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McKeague

et al. 2020

Chem. Soc. Rev.

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Section: Methylating Agents (O 6 -Methylguanine)mentioning

confidence: 96%

Next-generation DNA damage sequencing

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McKeague

et al. 2020

Chem. Soc. Rev.

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“…Adduct-directed nucleoside probes have also been exploited to probe the catalytic center of enzymes involved in repair and bypass of O 6 -alkylG adducts, e.g. TLS Pols or alkylguanine alkyltransferase . Translesion DNA synthesis continues DNA replication in the face of DNA damage, but the low fidelity of TLS Pols is responsible for many mutations observed in cancer .…”

Section: Synthetic Nucleosides As Probes To Investigate Translesion D...mentioning

confidence: 57%

“…TLS Pols or alkylguanine alkyltransferase. 28 Translesion DNA synthesis continues DNA replication in the face of DNA damage, but the low fidelity of TLS Pols is responsible for many mutations observed in cancer. 3 Therefore, elucidating the mechanisms of DNA adduct bypass by TLS Pols is crucial to understand DNA damage-induced mutagenesis.…”

Section: Investigate Translesion Dna Synthesismentioning

confidence: 99%

DNA Adduct-Directed Synthetic Nucleosides

et al. 2019

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Conspectus Chemical damage to DNA is a key initiator of adverse biological consequences due to disruption of the faithful reading of the genetic code. For example, O 6-alkylguanine (O 6-alkylG) DNA adducts are strongly miscoding during DNA replication when the damaged nucleobase is a template for polymerase-mediated translesion DNA synthesis. Thus, mutations derived from O 6-alkylG adducts can have severe adverse effects on protein translation and function and are an early event in the initiation of carcinogenesis. However, the low abundance of these adducts places significant limitations on our ability to relate their presence and biological influences with resultant mutations or disease risk. As a consequence, there is a critical need for novel tools to detect and study the biological role of alkylation adducts. Incorporating DNA bases with altered structures that are derived synthetically is a strategy that has been used widely to interrogate biological processes involving DNA. Such synthetic nucleosides have contributed to our understanding of DNA structure, DNA polymerase (Pol) and repair enzyme function, and to the expansion of the genetic alphabet. This Account describes our efforts toward creating and applying synthetic nucleosides directed at DNA adducts. We synthesized a variety of nucleosides with altered base structures that complement the altered hydrogen bonding capacity and hydrophilicity of O 6-alkylG adducts. The heterocyclic perimidinone-derived nucleoside Per was the first of such adduct-directed synthetic nucleosides; it specifically stabilized O 6-benzylguanine (O 6-BnG) in a DNA duplex. Structural variants of Per were used to determine hydrogen bonding and base-stacking contributions to DNA duplex stability in templates containing O 6-BnG as well as O 6-methylguanine (O 6-MeG) adducts. We created synthetic probes able to stabilize damaged over undamaged templates and established how altered hydrogen bonding or base-stacking properties impact DNA duplex stability as a function of adduct structures. This knowledge was then applied to devise a hybridization-based detection strategy involving gold nanoparticles that distinguish damaged from undamaged DNA by colorimetric changes. Furthermore, synthetic nucleosides were used as mechanistic tools to understand chemical determinants such as hydrogen bonding, π-stacking, and size and shape deviations that impact the efficiency and fidelity of DNA adduct bypass by DNA Pols. Finally, we reported the first example of amplifying alkylated DNA, accomplished by combining an engineered polymerase and synthetic triphosphate for which incorporation is templated by a DNA adduct. The presence of the synthetic nucleoside in amplicons could serve as a marker for the presence and location of DNA damage at low levels in DNA strands. Adduct-directed synthetic nucleosides have opened new concepts to interrogate the levels, locations, and biological influences of DNA alkylation.

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