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Structural Insight into the Specific DNA Template Binding to DnaG primase in Bacteria
Yingqin Zhou1,2,3, Hao Luo1,2,3, Zhongchuan Liu1,2
1Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China.
Scientific Reports
|April 8, 2017
Summary
Bacterial primase (DnaG) initiates DNA replication by synthesizing RNA primers. This study reveals how BsuDnaG uses its zinc binding domain and an L-shaped surface to recognize specific DNA sequences, clarifying initiation site selection.
Area of Science:
- Molecular Biology
- Structural Biology
- Biochemistry
Background:
- Bacterial primase synthesizes RNA primers for DNA replication.
- The mechanism of primase's specific initiation site recognition is not fully understood.
Purpose of the Study:
- To characterize Bacillus subtilis DnaG primase (BsuDnaG).
- To determine the crystal structure of the BsuDnaG RNA polymerase domain (RPD).
- To elucidate the mechanism of primase-template DNA interaction and initiation site recognition.
Main Methods:
- X-ray crystallography to determine the RPD structure.
- Biochemical assays to study primase-template interactions.
- Structural comparisons of DnaG primases.
Main Results:
- The crystal structure of BsuDnaG RPD was determined.
- The tethered zinc binding domain is crucial for primase-template sequence interactions.
- An L-shaped surface was identified as the single-stranded DNA template binding site.
- The intrinsic flexibility of DnaG primases was highlighted.
Conclusions:
- The zinc binding domain and L-shaped surface mediate specific template recognition.
- Primase flexibility likely facilitates interactions within the replisome.
- This study provides insights into the DnaG-mediated DNA replication initiation mechanism.