Cryptic Protein Interactions Regulate DNA Replication Initiation

Overview

Journal Mol Microbiol

Specialties Microbiology
Molecular Biology

Date 2018 Oct 5

PMID 30285297

Citations 9

Authors

Lindsay A Matthews

Lyle A Simmons

Affiliations

Soon will be listed here.

Abstract

DNA replication is a fundamental biological process that is tightly regulated in all cells. In bacteria, DnaA controls when and where replication begins by building a step-wise complex that loads the replicative helicase onto chromosomal DNA. In many low-GC Gram-positive species, DnaA recruits the DnaD and DnaB proteins to function as adaptors to assist in helicase loading. How DnaA, its adaptors and the helicase form a complex at the origin is unclear. We addressed this question using the bacterial two-hybrid assay to determine how the initiation proteins from Bacillus subtilis interact with each other. We show that cryptic interaction sites play a key role in this process and we map these regions for the entire pathway. In addition, we found that the SirA regulator that blocks initiation in sporulating cells binds to a surface on DnaA that overlaps with DnaD. The interaction between DnaA and DnaD was also mapped to the same DnaA surface in the human pathogen Staphylococcus aureus, demonstrating the broad conservation of this surface. Therefore, our study has unveiled key protein interactions essential for initiation and our approach is widely applicable for mapping interactions in other signaling pathways that are governed by cryptic binding surfaces.

Citing Articles

Diverse Mechanisms of Helicase Loading during DNA Replication Initiation in Bacteria.

Blaine H, Simmons L, Stallings C J Bacteriol. 2023; 205(4):e0048722.

PMID: 36877032 PMC: 10128896. DOI: 10.1128/jb.00487-22.

SirA inhibits the essential DnaA:DnaD interaction to block helicase recruitment during Bacillus subtilis sporulation.

Winterhalter C, Stevens D, Fenyk S, Pelliciari S, Marchand E, Soultanas P Nucleic Acids Res. 2022; 51(9):4302-4321.

PMID: 36416272 PMC: 10201431. DOI: 10.1093/nar/gkac1060.

Structure and kinase activity of bacterial cell cycle regulator CcrZ.

Wozniak K, Burby P, Nandakumar J, Simmons L PLoS Genet. 2022; 18(5):e1010196.

PMID: 35576203 PMC: 9135335. DOI: 10.1371/journal.pgen.1010196.

DNA damage checkpoint activation affects peptidoglycan synthesis and late divisome components in Bacillus subtilis.

Masser E, Burby P, Hawkins W, Gustafson B, Lenhart J, Simmons L Mol Microbiol. 2021; 116(2):707-722.

PMID: 34097787 PMC: 8600973. DOI: 10.1111/mmi.14765.

Molecular insights into replication initiation in a multipartite genome harboring bacterium Deinococcus radiodurans.

Maurya G, Chaudhary R, Pandey N, Misra H J Biol Chem. 2021; 296:100451.

PMID: 33626388 PMC: 7988490. DOI: 10.1016/j.jbc.2021.100451.

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