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Ewing Sarcoma Related protein 1 recognizes R-loops by binding DNA forks
Michelle A Lay1,2,3, Valery F Thompson1,2, Ajibola D Adelakun1,2,4
1Department of Pharmacology, University of Arizona, Tucson, AZ 85724, USA.
Biorxiv : the Preprint Server for Biology
|January 31, 2024
Summary
Ewing Sarcoma Related protein 1 (EWSR1) binds R-loops, structures linked to genomic stress. Its Zinc-finger and RNA recognition motifs enable specific DNA fork recognition, crucial for R-loop regulation.
Area of Science:
- Molecular Biology
- Genomics
- Biochemistry
Background:
- EWSR1 (Ewing Sarcoma Related protein 1) is a ubiquitous RNA-binding protein involved in RNA processing.
- Dysregulation of EWSR1 is linked to increased R-loops, which cause genomic and transcription stress.
- R-loops are nucleic acid structures comprising a DNA:RNA hybrid and a displaced single-stranded DNA.
Approach:
- Investigated the specific mechanism by which EWSR1 recognizes R-loop structures.
- Utilized electrophoretic mobility shift assays (EMSA) to assess EWSR1 binding affinity to R-loop components.
- Focused on identifying the specific domains within EWSR1 responsible for R-loop recognition and specificity.
Key Points:
- EWSR1 exhibits high-affinity binding to substrates mimicking R-loop structures.
- Specificity of EWSR1 binding is localized to the DNA fork region of R-loops.
- The Zinc-finger (ZnF) and RNA recognition motif (RRM) domains, along with flanking RGG domains, are critical for EWSR1's binding affinity and specificity.
Conclusions:
- EWSR1's ZnF and RGG domains mediate high-affinity binding to R-loop DNA forks.
- EWSR1's RRM and RGG domains enhance specificity for R-loop structures.
- This domain-based recognition mechanism explains EWSR1's role in regulating R-loops across various cellular contexts.
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