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Force-Dependent Intercalative Bulky DNA Adduct Formation Detected by Single-Molecule Stretching
Yajun Liu1, Yufeng Pei1, Jingjing Xu1,2
1Hubei Key Laboratory of Natural Medicinal Chemistry and Resource Evaluation, School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Analytical Chemistry
|September 21, 2022
Summary
This study introduces a single-molecule stretching assay to analyze DNA adducts, revealing how DNA adduct formation correlates with intercalation and is enhanced by external forces.
Area of Science:
- Biochemistry
- Molecular Biology
- Chemical Biology
Background:
- Understanding DNA adducts is crucial for assessing drug/carcinogen toxicity.
- Current biochemical methods struggle with unstable adducts and topology analysis.
- A need exists for precise methods to quantify adduct formation and properties.
Purpose of the Study:
- To develop and apply a single-molecule stretching assay for characterizing intercalative DNA adducts.
- To investigate the kinetics and mechanical properties of various DNA adducts.
- To explore the influence of external forces on adduct formation.
Main Methods:
- Utilized single-molecule stretching assay to measure DNA elongation caused by adducts.
- Analyzed formaldehyde-mediated anthracycline-DNA adducts.
- Studied UV light-catalyzed psoralen-DNA adducts.
- Investigated liver S9 fraction-catalyzed aflatoxin B1-DNA adducts.
Main Results:
- Demonstrated correlation between adduct formation and noncovalent intercalation binding.
- Quantified the number and formation kinetics of intercalative DNA adducts.
- Showed that external forces significantly increase DNA adduct formation (1.8- to 5.3-fold).
Conclusions:
- Single-molecule stretching assay provides a powerful tool for studying DNA adducts.
- Adduct formation is intrinsically linked to intercalation efficiency.
- Mechanical forces can modulate the formation of DNA adducts, impacting their biological effects.
