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Updated: Jan 18, 2026

Analyzing and Building Nucleic Acid Structures with 3DNA
Published on: April 26, 2013
A single helicase-binding domain of DnaG couples with hexameric helicase DnaB in Bacillus stearothermophilus
Hao Luo1,2, Wenlin Liu1,2, Yingqin Zhou1
1Agricultural Microbial Agents Key Laboratory of Sichuan Province, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610213, China.
Abstract:
In bacterial DNA replication, helicase DnaB and primase DnaG form the primosome. Helicase DnaB unwinds double-stranded DNA (dsDNA) to provide templates for DNA polymerase, whereas primase DnaG supplies RNA primers to DNA polymerase for the synthesis of Okazaki fragments. How primase DnaG coordinates with helicase DnaB at the DNA replication fork remains unclear. In this study, the interactions between the helicase-binding domain of DnaG (DnaG (HBD)) and DnaB hexamer were studied. A stable ternary complex of DnaB6/dT16/DnaG(HBD) from Bacillus stearothermophilus was prepared and the homogeneity of the DnaB6/dT16/DnaG(HBD) complex was verified by dynamic light scattering. The stoichiometry of DnaG(HBD) to process DnaB6 was investigated by isothermal titration calorimetry. The results show that a single primase DnaG binds to DnaB6 in the presence of single-stranded DNA. Based on these results, a model is proposed to explain how the primase DnaG couples with the processing DnaB6 helicase during the Okazaki fragment synthesis cycle. These findings provide valuable insights into the coupling between dsDNA unwinding and RNA primer synthesis in DNA replication.
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