Cellular Assays to Study the Functional Importance of Human DNA Repair Helicases.
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.
['Awate|Sanket|S|', 'Dhar|Srijita|S|', 'Sommers|Joshua A|JA|', 'Brosh|Robert M|RM|']
D017209:Apoptosis D001681:Biological Assay D002452:Cell Count D045744:Cell Line, Tumor D049109:Cell Proliferation D004265:DNA Helicases D004260:DNA Repair D057075:Enzyme Assays D042822:Genomic Instability D006801:Humans D008856:Microscopy, Fluorescence
DNA repair Helicase DNA damage Genomic stability Genetic disease Replication stress