Biochemical methods to monitor loading and activation of hexameric helicases

Amy J. Fernandez, James M. Berger

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Ring-shaped hexameric helicases are an essential class of enzymes that unwind duplex nucleic acids to support a variety of cellular processes. Because of their critical roles in cells, hexameric helicase dysfunction has been linked to DNA damage and genomic instability. Biochemical characterization of hexameric helicase activity and regulation in vitro is necessary for understanding enzyme function and aiding drug discovery efforts. In this chapter, we describe protocols for characterizing mechanisms of helicase loading, activation, and unwinding using the model replicative hexameric DnaB helicase and its cognate DnaC loading factor from E. coli.

Original languageEnglish (US)
Title of host publicationHelicase Enzymes Part A
EditorsMichael A. Trakselis
PublisherAcademic Press Inc.
Pages143-152
Number of pages10
ISBN (Print)9780323914765
DOIs
StatePublished - Jan 2022

Publication series

NameMethods in Enzymology
Volume672
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • DNA unwinding
  • Helicase loading
  • Hexameric helicase

ASJC Scopus subject areas

  • Molecular Biology
  • Biochemistry

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