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Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry
Published on: January 17, 2025
BID binds to replication protein A and stimulates ATR function following replicative stress
Yang Liu1, Sivaraja Vaithiyalingam, Qiong Shi
1Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
Molecular and Cellular Biology
|August 24, 2011
Summary
BH3-interacting death domain agonist (BID) protein enhances DNA damage response by linking replication protein A (RPA) to sensor complexes. This BID-RPA interaction is crucial for ATR-ATRIP recruitment and ATR kinase function.
Area of Science:
- Molecular Biology
- Cellular Biology
- Biochemistry
Background:
- The BH3-interacting death domain agonist (BID) protein plays a role in apoptosis and DNA damage response.
- Replicative stress triggers the association of BID with DNA damage sensor proteins like replication protein A (RPA), ataxia telangiectasia and Rad3 related (ATR), and ATR-interacting protein (ATRIP).
Purpose of the Study:
- To elucidate the mechanism by which BID facilitates the DNA damage response.
- To investigate the role of the BID-RPA interaction in the recruitment of ATR-ATRIP and ATR kinase activity.
Main Methods:
- Investigated protein-protein interactions using biochemical assays.
- Assessed DNA damage response by measuring CHK1 activation and DNA replication recovery after hydroxyurea (HU) treatment.
- Utilized disruption of BID-RPA interaction to study downstream effects.
Main Results:
- BID directly stimulates the association of RPA with DNA damage sensor complex components via interaction with the RPA70 subunit.
- Disruption of the BID-RPA interaction impairs ATR-ATRIP chromatin association and ATR kinase function.
- BID-RPA interaction is shown to promote ATR-ATRIP association with the DNA damage sensor complex.
Conclusions:
- BID acts as a crucial mediator in the DNA damage response by bridging RPA to the sensor complex.
- The BID-RPA interaction is essential for efficient ATR-ATRIP recruitment and subsequent ATR kinase activation.
- A model is proposed where BID binding to RPA facilitates ATR-ATRIP recruitment and stabilization at DNA damage sites.
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