Related Experiment Video
Updated: May 31, 2026
05:37
Single-Molecule Fluorescence Visualization of DNA Polymerase Dynamics at G-Quadruplexes
Published on: April 4, 2025
The Rev1 translesion synthesis polymerase has multiple distinct DNA binding modes
Frederik H de Groote1, Jacob G Jansen, Yuji Masuda
1Department of Protein Chemistry, Leiden Institute of Chemistry, Gorlaeus Laboratory, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
DNA Repair
|July 15, 2011
Summary
Rev1, a DNA polymerase crucial for DNA repair, binds specifically to primer-template junctions. This binding, essential for translesion synthesis (TLS), involves its unique N-terminal BRCT region and an adjacent alpha-helix.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Rev1 is a eukaryotic DNA polymerase of the Y family.
- It plays a key role in translesion synthesis (TLS), a DNA damage tolerance pathway.
- Rev1 possesses a unique N-terminal BRCA1 C-terminal homology (BRCT) region.
Purpose of the Study:
- To investigate the DNA binding properties of yeast and mouse Rev1.
- To characterize the role of the BRCT region and adjacent elements in DNA binding.
Main Methods:
- Studied DNA binding properties of Rev1 from yeast and mouse.
- Analyzed the contribution of the N-terminal BRCT region, alpha-helix, and surrounding amino acids to DNA binding affinity.
Main Results:
- The BRCT region of Rev1 specifically binds to 5' phosphorylated, recessed primer-template junctions.
- This binding is dependent on an alpha-helix N-terminal to the BRCT domain and a 20-amino acid stretch preceding it.
- Rev1 also binds efficiently to recessed 3' primer-template junctions.
Conclusions:
- Rev1 exhibits dual DNA binding characteristics at primer-template junctions.
- These properties support the proposed model of Rev1 recruitment at stalled replication forks during TLS.
More Related Videos
Related Concept Videos
02:10Translesion DNA Polymerases
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
03:01The Replisome
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
03:01The Replisome
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with the...
01:31Proofreading
Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore, it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase Enzyme
Errors During Replication are Corrected by the DNA Polymerase Enzyme
01:43Proofreading
Overview
02:31Homologous 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...