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Replication in Prokaryotes

Replication in Prokaryotes

Replication in Prokaryotes

Replication in Prokaryotes

The DNA Replication Fork

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork. Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...

The DNA Replication Fork

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork. Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...

Replication in Prokaryotes

Replication in Prokaryotes

DNA replication has three main steps: initiation, elongation, and termination. Replication in prokaryotes begins when initiator proteins bind to the single origin of replication (ori) on the cell's circular chromosome. Replication then proceeds around the entire circle of the chromosome in each direction from the two replication forks, resulting in two DNA molecules.
Many Proteins Work Together to Replicate the Chromosome
Replication is coordinated and carried out by a host of specialized...

Replication in Eukaryotes

Replication in Eukaryotes

In eukaryotic cells, DNA replication is highly conserved and tightly regulated. Multiple linear chromosomes must be duplicated with high fidelity before cell division, so there are many proteins that fulfill specialized roles in the replication process. Replication occurs in three phases: initiation, elongation, and termination, and ends with two complete sets of chromosomes in the nucleus.
Many Proteins Orchestrate Replication at the Origin
Eukaryotic replication follows many of the same...

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Related Experiment Video

Updated: Jul 5, 2026

G2-seq: A High Throughput Sequencing-based Technique for Identifying Late Replicating Regions of the Genome

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Conformational changes in a replication origin induced by an initiator protein.

S Mukherjee, I Patel, D Bastia

|November 1, 1985

Summary

The R6K plasmid initiator protein binds to replication origins, causing DNA bending and unwinding. This interaction influences DNA structure and topoisomerase activity, crucial for replication initiation.

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Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

Strand-Specific Analysis of Proteins at Replicating DNA Strands by Enrichment and Sequencing of Protein-Associated Nascent DNA Method

Published on: May 2, 2025

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

Plasmid R6K relies on a replication initiator protein for DNA replication.
Replication origins contain essential direct repeat sequences for initiator binding.

Purpose of the Study:

To investigate the structural changes induced by the R6K initiator protein at replication origins.
To understand the mechanism of replication initiation and regulation.

Main Methods:

DNA-protein binding assays using plasmid R6K origins.
Analysis of DNA bending and unwinding upon initiator binding.
Assays for topoisomerase activity in the presence of the initiator protein.

Main Results:

Initiator protein binding induced significant DNA bending and unwinding at the gamma replication origin.
Distamycin inhibited initiator binding to the direct repeats.
Initiator protein enhanced topoisomerase-induced catenation of origin-containing DNA.

Conclusions:

The R6K initiator protein causes substantial alterations in the secondary and tertiary structures of replication origins.
These structural changes are critical for initiating DNA replication and are modulated by other factors.