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Biochemical and cell biological assays to identify and characterize DNA helicase inhibitors
Taraswi Banerjee1, Monika Aggarwal2, Joshua A Sommers1
1Laboratory of Molecular Gerontology, National Institute on Aging, National Institutes of Health, NIH Biomedical Research Center, 251 Bayview Blvd, Baltimore, MD 21224, USA.
Abstract:
The growing number of DNA helicases implicated in hereditary disorders and cancer indicates that this particular class of enzymes plays key roles in genomic stability and cellular homeostasis. Indeed, a large body of work has provided molecular and cellular evidence that helicases act upon a variety of nucleic acid substrates and interact with numerous proteins to enact their functions in replication, DNA repair, recombination, and transcription. Understanding how helicases operate in unique and overlapping pathways is a great challenge to researchers. In this review, we describe a series of experimental approaches and methodologies to identify and characterize DNA helicase inhibitors which collectively provide an alternative and useful strategy to explore their biological significance in cell-based systems. These procedures were used in the discovery of biologically active compounds that inhibited the DNA unwinding function catalyzed by the human WRN helicase-nuclease defective in the premature aging disorder Werner syndrome. We describe in vitro and in vivo experimental approaches to characterize helicase inhibitors with WRN as the model, anticipating that these approaches may be extrapolated to other DNA helicases, particularly those implicated in DNA repair and/or the replication stress response.
Insights
Researchers developed new methods to find and study DNA helicase inhibitors. These tools helped discover compounds that block the WRN helicase, offering a strategy to explore helicase functions in diseases like Werner syndrome.
Area of Science:
- Biochemistry and Molecular Biology
- Genetics and Genomics
Background:
- DNA helicases are crucial for genomic stability and cellular homeostasis.
- Dysfunctional DNA helicases are linked to hereditary disorders and cancer.
- Understanding diverse helicase functions in replication, repair, and transcription is challenging.
Purpose of the Study:
- To present experimental approaches for identifying and characterizing DNA helicase inhibitors.
- To explore the biological significance of helicases using inhibitor-based strategies.
- To establish a model for studying helicase inhibitors, exemplified by the WRN helicase.
Main Methods:
- Described a series of experimental methodologies for DNA helicase inhibitor discovery and characterization.
- Utilized in vitro and in vivo approaches to study inhibitor activity.
- Focused on the human WRN helicase-nuclease, defective in Werner syndrome, as a model system.
Main Results:
- Successfully identified biologically active compounds inhibiting DNA unwinding by the WRN helicase.
- Demonstrated the utility of the developed methods in discovering functional helicase inhibitors.
- Provided a framework for extrapolating these methods to other DNA helicases.
Conclusions:
- Experimental approaches for DNA helicase inhibitor characterization are effective.
- Inhibitor-based studies offer a valuable strategy to understand helicase roles in cellular processes.
- The described methods can be applied to investigate DNA helicases involved in DNA repair and replication stress response.
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