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Updated: Oct 7, 2025
08:56
In Situ Detection of Ribonucleoprotein Complex Assembly in the C. elegans Germline using Proximity Ligation Assay
Published on: May 5, 2020
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Proximity labeling identifies a repertoire of site-specific R-loop modulators
Qingqing Yan1,2, Phillip Wulfridge1,2, John Doherty1,2
1Gene Expression and Regulation Program, The Wistar Institute, Philadelphia, PA, 19104, USA.
Nature Communications
|January 11, 2022
Summary
R-loops, linked to genome instability, are regulated by proteins like ATRX and ADNP. ADNP
Area of Science:
- Molecular Biology
- Genetics
- Neuroscience
Background:
- R-loops are three-stranded nucleic acid structures implicated in genome instability, accumulating in neurological diseases and cancers.
- Understanding R-loop regulation is crucial for addressing associated pathologies.
Purpose of the Study:
- To identify and characterize proteins that regulate R-loops in vivo.
- To investigate the role of activity-dependent neuroprotective protein (ADNP) in R-loop resolution and its implications for neuronal function and disease.
Main Methods:
- Utilized a proximity-dependent labeling system for proteomic screening.
- Performed in vitro and in vivo experiments to assess R-loop resolution by ADNP.
- Analyzed patient-derived induced pluripotent stem cells with ADNP mutations.
Main Results:
- Identified distinct protein classes regulating R-loops, including ATRX and zinc finger/homeodomain proteins.
- ADNP was found to resolve R-loops in vitro and suppress them in vivo at its genomic targets.
- ADNP mutations compromised R-loop resolution, leading to R-loop accumulation and impaired neuronal differentiation, observed in patient-derived cells.
Conclusions:
- ADNP plays a critical role in resolving R-loops, essential for normal neuronal function.
- Dysregulation of ADNP-mediated R-loop resolution is linked to ADNP syndrome and potentially other developmental disorders and cancers.
- Zinc finger and homeodomain proteins are broadly implicated in R-loop regulation.
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