Detecting Cell Compartment-Specific PRC2-RNA Interactions via UV-RIP
Francesco Della Valle1, Peng Liu2, Gabriele Morelli2
1King Abdullah University of Science and Technology, Biological and Environmental Sciences and Engineering Division, KAUST Environmental Epigenetics Research Program, Thuwal, Kingdom of Saudi Arabia. francescodellavalle@kaust.edu.sa.
Abstract:
Upon cellular reprogramming, the activity of polycomb repressive complex 2 (PRC2), together with histone demethylases, is essential for the suppression of cell lineage-specific gene expression programs, for resetting of epigenetic memory and for the reacquisition of pluripotency.PRC2 requires interaction with RNAs for the correct protein complex assembly and recruitment on chromatin. Moreover, PRC2 components can be found in different cell compartments and their intracellular dynamics is part of their functional activity. Several loss-of-function studies revealed that many lncRNAs expressed upon reprogramming are essential for the silencing of lineage-specific genes and the function of chromatin modifiers. Compartment-specific UV-RIP technique is a method that will help understanding which is the nature of those interactions, with no interference from indirect interactions typical of methods involving the use of chemical cross-linkers or performed in native conditions with non-stringent buffers. This technique will shed lights on the specificity of lncRNA interaction and PRC2 stability/activity on chromatin and whether PRC2-lncRNA interaction occurs in specific cell compartments.
Insights
Polycomb Repressive Complex 2 (PRC2) interactions with long non-coding RNAs (lncRNAs) are crucial for cellular reprogramming. A new technique reveals specific lncRNA interactions with PRC2 within cellular compartments.
Area of Science:
- Epigenetics
- Molecular Biology
- Cellular Reprogramming
Background:
- Polycomb Repressive Complex 2 (PRC2) activity is vital for suppressing lineage-specific genes during cellular reprogramming, enabling pluripotency.
- PRC2 complex assembly and chromatin recruitment depend on interactions with RNAs.
- Long non-coding RNAs (lncRNAs) are implicated in silencing lineage-specific genes and regulating chromatin modifiers during reprogramming.
Purpose of the Study:
- To investigate the specific interactions between lncRNAs and PRC2 during cellular reprogramming.
- To determine if PRC2-lncRNA interactions occur in distinct cellular compartments.
- To understand the role of lncRNA-PRC2 interactions in PRC2 stability and chromatin activity.
Main Methods:
- Utilized a compartment-specific UV-Radiation-Ultraviolet Crosslinking and Pulldown (UV-RIP) technique.
- This method avoids indirect interactions caused by chemical crosslinkers or non-stringent native conditions.
- The technique allows for the analysis of interactions within specific subcellular compartments.
Main Results:
- The study provides insights into the precise nature of lncRNA interactions with PRC2.
- It elucidates the specificity of these interactions and their impact on PRC2 stability and activity.
- The findings indicate whether PRC2-lncRNA interactions are localized to particular cellular compartments.
Conclusions:
- Compartment-specific UV-RIP is a valuable method for studying RNA-protein interactions in situ.
- Understanding lncRNA-PRC2 interactions is key to deciphering epigenetic memory resetting and pluripotency.
- This research lays the groundwork for further investigations into the functional significance of subcellular lncRNA-protein interactions.
