More Questions from DNA Structure and Replication

At the E. coli oriC, the initial melting of about 40–45 base pairs requires which minimal set of factors?

Biochemistry DNA Structure and Replication Difficulty: Medium
Choose an option
  • A
    DNA + ATP + HU
  • B
    DNA + ATP + HU + DnaA
  • C
    DNA + ATP + HU + DnaB
  • D
    DNA + ATP + HU + DnaB + DnaA
  • E
    DNA + ATP + HU + DnaA + DNA polymerase III holoenzyme

Answer

Correct Answer: DNA + ATP + HU + DnaA

Explanation

Introduction / Context:Initiation at the E. coli origin (oriC) proceeds through ordered steps: origin recognition, DNA unwinding (melting), helicase loading, and replisome assembly. The question asks for the minimal factors needed for the initial 40–45 bp origin melting.

Given Data / Assumptions:

  • Organism: E. coli, origin oriC.
  • Proteins: DnaA (initiator), DnaB (helicase), HU (DNA-bending protein).
  • ATP is available for DnaA activity.

Concept / Approach:DnaA-ATP binds DnaA boxes within oriC, oligomerizes, and induces local strand opening in the AT-rich DNA unwinding element (DUE). HU facilitates bending and melting. DnaB helicase acts after melting, being loaded by DnaC.

Step-by-Step Solution:1) DnaA in the ATP-bound state recognizes oriC and multimerizes.2) HU stabilizes wrapped/bent DNA, lowering the energy barrier for unwinding.3) This DnaA-ATP + HU complex opens ~40–45 bp at the DUE.4) Only after opening is DnaB loaded (by DnaC) to expand the replication forks.

Verification / Alternative check:In vitro reconstitution shows origin opening by DnaA-ATP with architectural proteins like HU; helicase loading is a subsequent step, not a prerequisite for initial melting.

Why Other Options Are Wrong:Options lacking DnaA miss the initiator. Options including DnaB or polymerase III add factors needed after melting, not for the minimal opening step.

Common Pitfalls:Equating “melting” with full fork establishment. Helicase action extends unwinding but is not required for the initial 40–45 bp opening.

Final Answer:DNA + ATP + HU + DnaA

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