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DNA Repair pp 185-207 | Cite as

Cellular Assays to Study the Functional Importance of Human DNA Repair Helicases

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1999)

Abstract

DNA helicases represent a specialized class of enzymes that play crucial roles in the DNA damage response. Using the energy of nucleoside triphosphate binding and hydrolysis, helicases behave as molecular motors capable of efficiently disrupting the many noncovalent hydrogen bonds that stabilize DNA molecules with secondary structure. In addition to their importance in DNA damage sensing and signaling, DNA helicases facilitate specific steps in DNA repair mechanisms that require polynucleotide tract unwinding or resolution. Because they play fundamental roles in the DNA damage response and DNA repair, defects in helicases disrupt cellular homeostasis. Thus, helicase deficiency or inhibition may result in reduced cell proliferation and survival, apoptosis, DNA damage induction, defective localization of repair proteins to sites of genomic DNA damage, chromosomal instability, and defective DNA repair pathways such as homologous recombination of double-strand breaks. In this chapter, we will describe step-by-step protocols to assay the functional importance of human DNA repair helicases in genome stability and cellular homeostasis.

Key words

Helicase DNA repair Genomic stability DNA damage Replication stress Genetic disease 
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Notes

Acknowledgments

This work is supported by the National Institutes of Health, National Institute on Aging.

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratory of Molecular Gerontology, National Institute on Aging, NIHNIH Biomedical Research CenterBaltimoreUSA

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