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

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.

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 Correct Timing of dnaA Transcription and Initiation of DNA Replication Requires trans Translation

2009 ◽  
Vol 191 (13) ◽  
pp. 4268-4275 ◽  
Author(s):  
Lin Cheng ◽  
Kenneth C. Keiler
Keyword(s):  
Gene Expression ◽  
Dna Replication ◽  
Caulobacter Crescentus ◽  
Replication Initiation ◽  
Proximal Promoter ◽  
Dna Replication Initiation ◽  
Proliferation And Differentiation ◽  
Initiation Of Dna Replication ◽  
Protein Tagging

ABSTRACT The trans translation pathway for protein tagging and ribosome release has been found in all bacteria and is required for proliferation and differentiation in many systems. Caulobacter crescentus mutants that lack the trans translation pathway have a defect in the cell cycle and do not initiate DNA replication at the correct time. To determine the molecular basis for this phenotype, effects on events known to be important for initiation of DNA replication were investigated. In the absence of trans translation, transcription from the dnaA promoter and an origin-proximal promoter involved in replication initiation is delayed. Characterization of the dnaA promoter revealed two cis-acting elements that have dramatic effects on dnaA gene expression. A 5′ leader sequence in dnaA mRNA represses gene expression by >15-fold but does not affect the timing of dnaA expression. The second cis-acting element, a sequence upstream of the −35 region, affects both the amount of dnaA transcription and the timing of transcription in response to trans translation. Mutations in this promoter element eliminate the transcription delay and partially suppress the DNA replication phenotype in mutants lacking trans translation activity. These results suggest that the trans translation capacity of the cell is sensed through the dnaA promoter to control the timing of DNA replication initiation.

A Novel Fluorescence-Based Screen for Inhibitors of the Initiation of DNA Replication in Bacteria

2019 ◽  
Vol 16 (3) ◽  
pp. 272-277 ◽  
Author(s):  
Rasmus N. Klitgaard ◽  
Anders Løbner-Olesen
Keyword(s):  
Dna Replication ◽  
High Throughput ◽  
Cyclic Peptide ◽  
Replication Initiation ◽  
False Positive Result ◽  
Over Expression ◽  
Initiator Protein ◽  
Dna Replication Initiation ◽  
Cellular Processes ◽  
Initiation Of Dna Replication

Background:One of many strategies to overcome antibiotic resistance is the discovery of compounds targeting cellular processes, which have not yet been exploited.Materials and Methods:Using various genetic tools, we constructed a novel high throughput, cellbased, fluorescence screen for inhibitors of chromosome replication initiation in bacteria.Results:The screen was validated by expression of an intra-cellular cyclic peptide interfering with the initiator protein DnaA and by over-expression of the negative initiation regulator SeqA. We also demonstrated that neither tetracycline nor ciprofloxacin triggers a false positive result. Finally, 400 extracts isolated mainly from filamentous actinomycetes were subjected to the screen.Conclusion:We concluded that the presented screen is applicable for identifying putative inhibitors of DNA replication initiation in a high throughput setup.

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