Positive and negative control of helicase recruitment at a bacterial chromosome origin

2021 ◽  
Author(s):  
Charles Winterhalter ◽  
Daniel Stevens ◽  
Stepan Fenyk ◽  
Simone Pelliciari ◽  
Elie Marchand ◽  
...  
Keyword(s):  
Dna Replication ◽  
Model Organism ◽  
Negative Control ◽  
Replication Initiation ◽  
Bacterial Dna ◽  
Dna Replication Initiation ◽  
Recognition Element ◽  
Helicase Loading ◽  
Protein Interfaces

The mechanisms responsible for helicase loading during the initiation of chromosome replication in bacteria are unclear. Here we report both a positive and a negative mechanism for directing helicase recruitment in the model organism Bacillus subtilis. Systematic mutagenesis of the essential replication initiation gene dnaD and characterization of DnaD variants revealed protein interfaces required for interacting with the master initiator DnaA and with a specific single-stranded DNA (ssDNA) sequence located in the chromosome origin (DnaD Recognition Element, DRE). We propose that the location of the DRE within the replication origin orchestrates recruitment of helicase to achieve bidirectional DNA replication. We also report that the developmentally expressed repressor of DNA replication initiation, SirA, acts by blocking the interaction of DnaD with DnaA, thereby inhibiting helicase recruitment to the origin. These findings significantly advance our mechanistic understanding of helicase recruitment and regulation during bacterial DNA replication initiation. Because DnaD is essential for the viability of clinically relevant Gram-positive pathogens, DnaD is an attractive target for drug development.

scholarly journals Near-Atomic Structural Model for Bacterial DNA Replication Initiation Complex and its Functional Insights

2017 ◽  
Vol 112 (3) ◽  
pp. 512a
Author(s):  
Masahiro Shimizu ◽  
Yasunori Noguchi ◽  
Yukari Sakiyama ◽  
Hironori Kawakami ◽  
Tsutomu Katayama ◽  
...  
Keyword(s):  
Dna Replication ◽  
Structural Model ◽  
Replication Initiation ◽  
Initiation Complex ◽  
Bacterial Dna ◽  
Dna Replication Initiation ◽  
Bacterial Dna Replication

Characterization of DNA synthesis and chloroplast DNA replication initiation in a Petunia hybrida chloroplast lysate system

1987 ◽  
Vol 12 (5) ◽  
pp. 377-386 ◽  
Author(s):  
Jan M. de Haas ◽  
Ad J. Kool ◽  
Nico Overbeeke ◽  
Wieger van Brug ◽  
H. John ◽  
...  
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Chloroplast Dna ◽  
Petunia Hybrida ◽  
Replication Initiation ◽  
Dna Replication Initiation ◽  
Chloroplast Dna Replication

Cloning and characterization of Paramecium mitochondrial DNA replication initiation regions

Gene ◽  
1980 ◽  
Vol 11 (1-2) ◽  
pp. 43-52 ◽  
Author(s):  
Arthur E. Pritchard ◽  
Lynne M. Herron ◽  
Donald J. Cummings
Keyword(s):  
Mitochondrial Dna ◽  
Dna Replication ◽  
Replication Initiation ◽  
Dna Replication Initiation ◽  
Mitochondrial Dna Replication

scholarly journals DNA replication initiation in Bacillus subtilis: structural and functional characterization of the essential DnaA–DnaD interaction

2018 ◽  
Vol 47 (4) ◽  
pp. 2101-2112 ◽  
Author(s):  
Eleyna Martin ◽  
Huw E L Williams ◽  
Matthaios Pitoulias ◽  
Daniel Stevens ◽  
Charles Winterhalter ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
Dna Replication ◽  
Functional Characterization ◽  
Replication Initiation ◽  
Dna Replication Initiation

scholarly journals Effects of oriC relocation on control of replication initiation in Bacillus subtilis

Microbiology ◽  
2009 ◽  
Vol 155 (9) ◽  
pp. 3070-3082 ◽  
Author(s):  
Shigeki Moriya ◽  
Yoshikazu Kawai ◽  
Sakiko Kaji ◽  
Adrian Smith ◽  
Elizabeth J. Harry ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Bacillus Subtilis ◽  
Dna Replication ◽  
Negative Control ◽  
Replication Initiation ◽  
Control Mechanisms ◽  
Bacterial Cells ◽  
Wild Type ◽  
Dna Replication Initiation ◽  
In The Wild

In bacteria, DNA replication initiation is tightly regulated in order to coordinate chromosome replication with cell growth. In Escherichia coli, positive factors and negative regulatory mechanisms playing important roles in the strict control of DNA replication initiation have been reported. However, it remains unclear how bacterial cells recognize the right time for replication initiation during the cell cycle. In the Gram-positive bacterium Bacillus subtilis, much less is known about the regulation of replication initiation, specifically, regarding negative control mechanisms which ensure replication initiation only once per cell cycle. Here we report that replication initiation was greatly enhanced in strains that had the origin of replication (oriC) relocated to various loci on the chromosome. When oriC was relocated to new loci further than 250 kb counterclockwise from the native locus, replication initiation became asynchronous and earlier than in the wild-type cells. In two oriC-relocated strains (oriC at argG or pnbA, 25 ° or 30 ° on the 36 ° chromosome map, respectively), DnaA levels were higher than in the wild-type but not enough to cause earlier initiation of replication. Our results suggest that the initiation capacity of replication is accumulated well before the actual time of initiation, and its release may be suppressed by a unique DNA structure formed near the native oriC locus.

scholarly journals Interactions between helicase and primase are crucial for DNA replication in the enteropathogen Clostridium difficile

2016 ◽  
Author(s):  
Erika van Eijk ◽  
Vasileios Paschalis ◽  
Matthew Green ◽  
Annemieke H. Friggen ◽  
Marilynn A. Larson ◽  
...  
Keyword(s):  
Dna Replication ◽  
Clostridium Difficile ◽  
De Novo ◽  
Model Organism ◽  
Replication Initiation ◽  
Aquifex Aeolicus ◽  
Dna Replication Initiation ◽  
Domains Of Life ◽  
Micro Organisms

ABSTRACTDNA replication is an essential and conserved process in all domains of life and may serve as a target for the development of new antimicrobials. However, such developments are hindered by a limited understanding of DNA replication in pathogenic micro-organisms. Clostridium difficile is the main cause of health-care associated diarrhea and its DNA replication machinery is virtually uncharacterized. We identified the replicative helicase (CD3657), the helicase loader ATPase (CD3654) and primase (CD1454) of C. difficile and reconstitute helicase and primase activity in vitro. We demonstrate a direct and ATP-dependent interaction between the helicase loader and the helicase. We find that helicase activity is dependent on the presence of primase in vitro. The trinucleotide specificity of primase, which we find to be determined by a single lysine residue, is similar to extreme thermophile Aquifex aeolicus but the presence of helicase allows more efficient de novo synthesis of RNA primers from non-preferred trinucleotides. Thus, helicase-primase interactions constitute a crucial aspect of DNA replication initiation in C. difficile and helicase loading and activation in this organism differs critically from that of the Gram-positive model organism Bacillus subtilis.

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