Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Single-Strand DNA Binding Proteins

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...

Mismatch Repair

Mismatch Repair

Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...

Mismatch Repair

Mismatch Repair

Bacterial RNA Polymerase

Bacterial RNA Polymerase

Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...

Cooperative Binding of Transcription Regulators

Cooperative Binding of Transcription Regulators

Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome. Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form dimers that...

Conserved Binding Sites

Conserved Binding Sites

Many proteins’ biological role depends on their interactions with their ligands, small molecules that bind to specific locations on the protein known as ligand-binding sites. Ligand-binding sites are often conserved among homologous proteins as these sites are critical for protein function.
Binding sites are often located in large pockets, and if their location on a protein’s surface is unknown, it can be predicted using various approaches. The energetic method computationally analyses the...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

The transition from ITS-90 to primary thermometry above 1235 K.

Philosophical transactions. Series A, Mathematical, physical, and engineering sciences·2026

Same author

Future traceability of practical primary thermometry and self-validating thermometry.

Philosophical transactions. Series A, Mathematical, physical, and engineering sciences·2026

Same author

Novel primer-less amplification method for detection of RNA molecules.

Methods (San Diego, Calif.)·2025

Same author

A novel technology for margin extension and local control in breast conservation surgery: Saline-coupled intraoperative radiofrequency ablation (SIRA).

Surgical oncology·2025

Same author

S-phase checkpoint protects from aberrant replication fork processing and degradation.

Nucleic acids research·2025

Same author

Mouse PrimPol Outperforms Its Human Counterpart as a Robust DNA Primase.

International journal of molecular sciences·2025

Search research articles

Related Experiment Video

Updated: May 18, 2026

An Assay for Quantifying Protein-RNA Binding in Bacteria

An Assay for Quantifying Protein-RNA Binding in Bacteria

Published on: June 12, 2019

DNA-binding determinants promoting NHEJ by human Polμ.

Maria Jose Martin¹, Raquel Juarez, Luis Blanco

¹Department of Genome Dynamics and Function, Centro de Biologia Molecular Severo Ochoa (CSIC-UAM), 28049 Madrid, Spain.

Nucleic Acids Research

|October 5, 2012

Summary

Human Polµ is a unique DNA polymerase that intrinsically repairs DNA double-strand breaks (DSBs) via non-homologous end-joining (NHEJ). Specific residues and its BRCT domain are crucial for its DNA binding and repair functions.

More Related Videos

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Published on: November 10, 2016

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Published on: March 9, 2022

Related Experiment Videos

Last Updated: May 18, 2026

An Assay for Quantifying Protein-RNA Binding in Bacteria

An Assay for Quantifying Protein-RNA Binding in Bacteria

Published on: June 12, 2019

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks

Published on: November 10, 2016

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Deciphering Molecular Mechanism of Histone Assembly by DNA Curtain Technique

Published on: March 9, 2022

Area of Science:

Molecular Biology
Genetics
Biochemistry

Background:

Non-homologous end-joining (NHEJ) is the primary mechanism for repairing DNA double-strand breaks (DSBs) in eukaryotes.
DNA polymerases, including Polµ, are essential molecular tools in the NHEJ pathway.
Polµ exhibits a unique ability to synthesize DNA between non-complementary DNA ends during NHEJ.

Purpose of the Study:

To elucidate the intrinsic DNA-binding capabilities of human Polµ.
To identify the molecular determinants governing Polµ's specific function in NHEJ.
To investigate the role of the Polµ BRCT domain in DNA interaction and NHEJ enhancement.

Main Methods:

Electromobility shift assays (EMSA) to visualize DNA-protein interactions.
DNA footprinting assays to map protein-DNA binding sites.
Site-directed mutagenesis to assess the function of specific amino acid residues.

Main Results:

Human Polµ's capacity for DNA synthesis between non-complementary ends is an intrinsic property.
Stable DNA binding by Polµ requires a 5' phosphate group, orienting its catalytic domain.
Specific residues (Lys249, Arg253, Arg416) and the BRCT domain are critical for stabilizing DNA substrates and enhancing NHEJ.

Conclusions:

Polµ directly recognizes DNA ends via its 5' phosphate group, crucial for aligning substrates in NHEJ.
Key residues and the BRCT domain contribute to Polµ's DNA binding affinity and function in NHEJ.
These findings reveal novel aspects of Polµ's mechanism in DNA repair, highlighting its direct role in substrate processing.