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DNA Helicases

DNA Helicases

DNA unwinding helicase enzymes are a type of motor protein. Motor proteins can translocate along filaments or polymers using energy generated from ATP hydrolysis. Helicases are involved in all the important cellular processes where DNA unwinding is required, such as DNA replication, repair, recombination, and transcription. They are present in all living organisms, but vary in their structure, function, and mechanism of action. For example, in prokaryotes, DnaB helicase binds and translocates...

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

Gene Conversion

Gene Conversion

Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...

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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Related Experiment Video

Updated: Jun 15, 2026

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Published on: September 23, 2011

Mutational analysis of Bloom helicase.

¹Institut CURIE Recherche, Orsay, France.

Methods in Molecular Biology (Clifton, N.J.)

|March 13, 2010

Summary

This study investigates Bloom helicase mutations causing Bloom's syndrome, a cancer predisposition disorder. Researchers used structural modeling and biochemical methods to analyze DNA unwinding and binding activities, providing insights into disease mechanisms.

Area of Science:

Molecular Biology
Biochemistry
Genetics

Background:

DNA helicases are crucial motor proteins that unwind double-stranded DNA using ATP hydrolysis.
Essential for DNA replication, repair, recombination, and transcription.
Mutations in RecQ helicases, like Bloom helicase, cause human diseases including Bloom's syndrome, linked to cancer predisposition.

Purpose of the Study:

To elucidate the molecular basis of Bloom's syndrome pathology.
To understand the functional mechanisms of Bloom helicase and its disease-associated variants.
To establish methods for analyzing helicase activity relevant to human diseases.

Main Methods:

Structural modeling to predict mutation effects.
Site-directed mutagenesis to create specific Bloom helicase variants.

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Development of Targeting Induced Local Lesions IN Genomes (TILLING) Populations in Small Grain Crops by Ethyl Methanesulfonate Mutagenesis

Development of Targeting Induced Local Lesions IN Genomes (TILLING) Populations in Small Grain Crops by Ethyl Methanesulfonate Mutagenesis

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Last Updated: Jun 15, 2026

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae

Published on: September 23, 2011

Identifying Mutations by High Resolution Melting in a TILLING Population of Rice

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Development of Targeting Induced Local Lesions IN Genomes (TILLING) Populations in Small Grain Crops by Ethyl Methanesulfonate Mutagenesis

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Published on: July 16, 2019

Biochemical and biophysical assays to measure ATPase, DNA binding, DNA strand annealing, and DNA unwinding activities.

Main Results:

Detailed characterization of ATPase, ATP and DNA binding, DNA strand annealing, and DNA unwinding activities of wild-type and mutant Bloom helicase.
Identification of how specific missense mutations impact Bloom helicase function.
Establishment of a robust methodological framework for studying helicase dysfunction.

Conclusions:

The study provides critical insights into the molecular mechanisms underlying Bloom's syndrome.
The developed methods are valuable for studying other helicases and related genetic disorders.
Understanding these mechanisms can inform future therapeutic strategies for Bloom's syndrome and associated cancers.