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Related Concept Videos

Homologous Recombination02:31

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...
DNA Topoisomerases02:02

DNA Topoisomerases

Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types.  Type I...
Translesion DNA Polymerases02:10

Translesion 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...
Single-Strand DNA Binding Proteins01:03

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...
Nucleotide Excision Repair01:08

Nucleotide Excision Repair

Overview
Nucleotide Excision Repair01:38

Nucleotide Excision Repair

DNA Distortion and Damage
Cells are regularly exposed to mutagens—factors in the environment that can damage DNA and generate mutations. UV radiation is one of the most common mutagens and is estimated to introduce a significant number of changes in DNA. These include bends or kinks in the structure, which can block DNA replication or transcription. If these errors are not fixed, the damage can cause mutations, which in turn can result in cancer or disease depending on which sequences are...

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Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes
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Using Modified Synthetic Oligonucleotides to Assay Nucleic Acid-Metabolizing Enzymes

Published on: July 5, 2024

DNA ligases.

Gregory J S Lohman1, Stanley Tabor, Nicole M Nichols

  • 1New England Biolabs, Ipswich, Massachusetts, USA.

Current Protocols in Molecular Biology
|April 8, 2011
PubMed
Summary
This summary is machine-generated.

DNA ligases are crucial enzymes that join DNA fragments by forming phosphodiester bonds. This review details reaction conditions and applications for various DNA ligases, including T4, E. coli, and thermostable variants.

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Area of Science:

  • Molecular Biology
  • Enzymology
  • Biochemistry

Background:

  • DNA ligases catalyze the formation of phosphodiester bonds in duplex DNA.
  • This enzymatic activity is essential for sealing nicks and joining DNA fragments.
  • These enzymes play critical roles in DNA repair, replication, and recombinant DNA technology.

Purpose of the Study:

  • To provide a comprehensive overview of the DNA ligase enzyme family.
  • To describe reaction conditions and applications for key DNA ligases.
  • To highlight the differences in cofactor requirements, substrate specificity, and thermal stability among these enzymes.

Main Methods:

  • Review of established protocols and applications for DNA ligase enzymes.
  • Comparative analysis of T4 DNA ligase, E. coli DNA ligase, and thermostable DNA ligases.
  • Discussion of enzymatic mechanisms and reaction parameters.

Main Results:

  • DNA ligases effectively seal nicks and join DNA fragments with blunt or cohesive ends.
  • T4 DNA ligase, E. coli DNA ligase, and thermostable DNA ligases exhibit distinct properties.
  • Enzymes vary in their optimal reaction conditions, cofactor needs, and stability at different temperatures.

Conclusions:

  • DNA ligases are indispensable tools in molecular biology, nucleic acid research, and next-generation sequencing.
  • Understanding the specific characteristics of different DNA ligases allows for optimized application selection.
  • The diverse nature of DNA ligases provides flexibility for various experimental and biotechnological purposes.