DNA polymerase III holoenzyme

DNA polymerase III holoenzyme is the primary enzyme complex involved in DNA replication in Escherichia coli (E. coli) and many other bacteria. It is responsible for the majority of DNA synthesis during cellular replication. The holoenzyme is highly processive, meaning it can synthesize long stretches of DNA without dissociating from the template. Its activity is essential for the accuracy and speed of DNA replication, making it a critical component of the bacterial cell cycle.

Structure

The DNA polymerase III holoenzyme is a complex structure composed of multiple subunits, each with a specific function. The core of the enzyme consists of three subunits: α, ε, and θ. The α subunit (encoded by the dnaE gene) possesses the polymerase activity, synthesizing new DNA strands. The ε subunit (encoded by the dnaQ gene) has 3'→5' exonuclease activity, which allows for the correction of misincorporated nucleotides, enhancing the fidelity of DNA replication. The θ subunit (encoded by the holE gene) is thought to stabilize the ε subunit and enhance its exonuclease activity.

Surrounding the core is a clamp loader complex, composed of several subunits (τ, γ, δ, δ', and χψ), which is responsible for loading the β clamp (encoded by the dnaN gene) onto DNA. The β clamp is a dimer that encircles the DNA molecule, allowing the core enzyme to slide along the DNA without dissociating, thereby increasing the processivity of DNA synthesis.

Function

The primary function of DNA polymerase III holoenzyme is to replicate the bacterial genome accurately and efficiently. It achieves this through a series of coordinated steps:

1. **Initiation**: The process begins at the origin of replication, where the DNA is unwound by helicase, creating a replication fork. 2. **Loading of the β clamp**: The clamp loader complex opens the β clamp and places it around the DNA. 3. **Elongation**: The α subunit begins synthesizing the new DNA strand, using the original strand as a template. The β clamp moves with the polymerase, keeping it attached to the DNA. 4. **Proofreading**: The ε subunit proofreads the newly synthesized DNA, excising any incorrectly incorporated nucleotides. 5. **Termination**: Replication continues until the polymerase reaches the termination site, where it dissociates from the DNA.

Regulation

The activity of DNA polymerase III holoenzyme is tightly regulated to ensure that DNA replication occurs only once per cell cycle. This regulation involves the interaction of the holoenzyme with other proteins, such as DnaA (initiator protein), and the control of the availability of the β clamp and the clamp loader complex.

Clinical Significance

While DNA polymerase III holoenzyme is specific to bacteria, understanding its function and regulation can inform the development of antibacterial drugs. Inhibitors targeting the unique components of the bacterial replication machinery, such as the β clamp or the clamp loader complex, could provide new avenues for antibiotic development.

See Also

• DNA replication
• DNA polymerase
• Escherichia coli