DNA Replication—Opening the Replication Bubble Which protein class is primarily responsible for opening a replication bubble by separating parental DNA strands?
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ADNA helicases
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BSingle-stranded DNA-binding proteins (SSB/RPA)
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CDNA ligase
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DDNA topoisomerase
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ESliding clamp (PCNA/β clamp)
Answer
Correct Answer: DNA helicases
Explanation
Introduction / Context:DNA replication begins at origins where a replication bubble forms as parental strands are separated. Understanding which enzymes initiate strand separation versus those that stabilize or process DNA is fundamental to replication biology.
Given Data / Assumptions:
- Replication requires helicase, SSB, topoisomerase, primase, polymerase, clamp, and ligase.
- Question asks for the protein class that actively opens (unwinds) the duplex.
Concept / Approach:Helicases use NTP hydrolysis to translocate along DNA and separate complementary strands, generating single-stranded templates. SSB binds and stabilizes exposed single strands; topoisomerase relieves torsional stress; ligase seals nicks after synthesis; sliding clamp enhances polymerase processivity. Only helicase directly unwinds to create the bubble.
Step-by-Step Solution:
Step 1: Identify the motor protein that breaks base pairing: helicase.Step 2: Distinguish supportive roles: SSB prevents re-annealing; topoisomerase prevents supercoil buildup; ligase joins Okazaki fragments.Step 3: Conclude helicase is the principal opener of the bubble.Verification / Alternative check:Biochemical assays show helicase-dependent DNA unwinding; inhibition of helicase blocks bubble formation despite presence of SSB or topoisomerase.
Why Other Options Are Wrong:
- SSB stabilizes but does not unwind.
- Ligase seals nicks; no unwinding function.
- Topoisomerase resolves supercoils but does not separate base pairs.
- Sliding clamp increases polymerase processivity; not an unwinding factor.
Common Pitfalls:Attributing the opening to SSB because it binds single-stranded DNA; overlooking that SSB acts after helicase action.
Final Answer:DNA helicases