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Related Concept Videos

DNA Helicases00:55

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...
The Replisome03:01

The 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 Replisome03:01

The 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 DNA Replication Fork01:02

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...
The DNA Replication Fork01:02

The DNA Replication Fork

An organism’s genome needs to be duplicated in an efficient and error-free manner for its growth and survival. The replication fork is a Y-shaped active region where two strands of DNA are separated and replicated continuously. The coupling of DNA unzipping and complementary strand synthesis is a characteristic feature of a replication fork.   Organisms with small circular DNA, such as E. coli, often have a single origin of replication; therefore, they have only two replication forks, one in...
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...

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DNA structure specificity conferred on a replicative helicase by its loader.

Milind K Gupta1, John Atkinson, Peter McGlynn

  • 1School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, Scotland, United Kingdom.

The Journal of Biological Chemistry
|November 3, 2009
PubMed
Summary
This summary is machine-generated.

The accessory factor DnaC specifically inhibits the Escherichia coli DnaB helicase from translocating along double-stranded DNA. This regulation ensures DnaB activity is restricted to replication forks, maintaining genome stability.

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Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Replicative helicases unwind DNA during replication, interacting with both single-stranded and double-stranded DNA.
  • The Escherichia coli DnaB helicase unwinds DNA at the replication fork, but its DNA structure specificity is not fully understood.
  • Accessory factors like DnaC are crucial for helicase loading and function.

Purpose of the Study:

  • To investigate the role of DnaC in conferring DNA structure specificity to the DnaB helicase.
  • To determine how DnaC regulates DnaB translocation on different DNA structures.
  • To elucidate the mechanism by which DnaC modulates DnaB activity.

Main Methods:

  • In vitro assays measuring helicase translocation rates on single-stranded and double-stranded DNA.
  • Analysis of DnaB helicase activity in the presence of varying concentrations of DnaC.
  • Biochemical experiments to assess the requirement of ATP-bound DnaC for inhibition.

Main Results:

  • DnaC inhibits DnaB translocation on double-stranded DNA but not on single-stranded DNA when present in excess.
  • This inhibition is dependent on the ATP-bound form of DnaC.
  • DnaC's ATPase activity is stimulated during translocation on single-stranded DNA, relieving the inhibition on double-stranded DNA.

Conclusions:

  • DnaC confers DNA structure specificity to DnaB, restricting its activity to replication forks.
  • This regulatory mechanism ensures precise targeting of helicase activity, preventing off-target DNA interactions.
  • Analogous regulatory mechanisms may exist in other organisms for their replicative helicases.