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Multiple RPAs make WRN syndrome protein a superhelicase
Mina Lee1, Soochul Shin2, Heesoo Uhm2
1Center for Nano-Bio Measurement, Korea Research Institute of Standards and Science, Daejeon, Republic of Korea.
Replication protein A (RPA) transforms the Werner syndrome protein (WRN) from a weak DNA helicase into a superhelicase. Multiple RPA binding dramatically enhances WRN
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Werner syndrome protein (WRN) possesses helicase activity.
- Replication protein A (RPA) is known to stimulate WRN's helicase activity.
- The precise mechanism of RPA-mediated WRN stimulation remains unclear.
Purpose of the Study:
- To investigate the helicase activity of WRN.
- To elucidate the stimulation mechanism of WRN by RPA at the single-molecule level.
Main Methods:
- Single-molecule Förster Resonance Energy Transfer (smFRET).
- Magnetic tweezers assays.
Main Results:
- WRN alone exhibits limited, repetitive unwinding of DNA (tens of base pairs).
- Binding of multiple RPA molecules converts WRN into a highly processive superhelicase.
- Stimulated WRN unidirectionally unwinds over 1 kilobase of double-stranded DNA.
Conclusions:
- RPA binding fundamentally alters WRN's helicase activity and processivity.
- Cofactor binding can drastically change enzyme function and biological roles.
- This study provides a model for cofactor-induced enzyme modulation.
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