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Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

Homologous Recombination

Homologous Recombination

The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...

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Restarting Stalled Replication Forks

DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart, a...

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The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.

S-Cdk Initiates DNA Replication

S-Cdk Initiates DNA Replication

The cell cycle is a series of events leading to DNA duplication followed by the division of cell content to form two daughter cells. The cell cycle progresses in four stages—the cell increases in size (gap 1 or G1-phase), duplicates its DNA (synthesis or S-phase), prepares to divide (gap 2 or G2-phase), and divides (mitosis or M-phase).
Two states at the origin of replication
In eukaryotes, the initiation of replication occurs at many sites on the chromosomes, called the origins of replication.

Fixing Double-strand Breaks

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The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...

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Related Experiment Video

Updated: Jul 15, 2026

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Visualization of DNA Repair Proteins Interaction by Immunofluorescence

Published on: June 26, 2020

CST complex promotes second-strand synthesis in break-induced replication.

Pilendra Thakre¹, Jeff Wang^2,3, Liping Liu^2,3

¹Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, TX, USA.

Nature Structural & Molecular Biology

|July 13, 2026

Summary

The Cdc13-Stn1-Ten1 (CST) complex is crucial for DNA repair via break-induced replication (BIR). In yeast lacking CST, second-strand DNA synthesis during BIR is impaired, a finding conserved in human cells.

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Area of Science:

Molecular Biology
Genetics
DNA Repair Mechanisms

Background:

Break-induced replication (BIR) is a key pathway for repairing DNA breaks and maintaining telomeres.
While leading-strand synthesis in BIR is understood, the mechanism for second-strand synthesis remains unclear.

Purpose of the Study:

To elucidate the mechanism of second-strand DNA synthesis during BIR.
To investigate the role of the Cdc13-Stn1-Ten1 (CST) complex in BIR.

Main Methods:

Utilized yeast mutants lacking the CST complex.
Employed biochemical reconstitution with DNA substrates mimicking BIR intermediates.

Main Results:

Early BIR steps (resection, D-loop formation, first-strand synthesis) occur normally in CST-deficient yeast.
Second-strand synthesis is significantly impaired in the absence of the CST complex.
Yeast CST enhances DNA polymerase α-primase activity, promoting second-strand synthesis in BIR.

Conclusions:

The CST complex plays a critical, conserved role in facilitating second-strand DNA synthesis during BIR.
CST's function in BIR provides mechanistic insight into non-canonical DNA synthesis pathways.