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PUMILIO, but not RBMX, binding is required for regulation of genomic stability by noncoding RNA NORAD
Mahmoud M Elguindy1,2, Florian Kopp1, Mohammad Goodarzi3
1Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, United States.
Elife
|July 26, 2019
Summary
The long noncoding RNA NORAD maintains genome stability by regulating PUMILIO proteins in the cytoplasm. Interaction with RBMX in the nucleus is not essential for this function.
Area of Science:
- Molecular Biology
- Genetics
- Epigenetics
Background:
- NORAD is a conserved long noncoding RNA (lncRNA) crucial for mammalian genome stability.
- NORAD negatively regulates PUMILIO (PUM) proteins in the cytoplasm, and its loss or PUM hyperactivity causes genome instability and premature aging.
- A recent study suggested NORAD's role in genome stability involves interaction with RBMX in the nucleus.
Purpose of the Study:
- To investigate the distinct roles of NORAD's interactions with PUMILIO (PUM) and RBMX in maintaining genome stability in human cells.
- To clarify the subcellular localization of NORAD and its functional relevance.
Main Methods:
- RNA Fluorescence In Situ Hybridization (FISH) to determine NORAD localization.
- Cellular fractionation to separate cytoplasmic and nuclear fractions.
- Genetic rescue experiments to assess the necessity of PUM and RBMX binding for NORAD's function.
Main Results:
- NORAD predominantly localizes to the cytoplasm, irrespective of DNA damage.
- Genetic rescue experiments confirmed that PUM binding is essential for NORAD-mediated genome stability.
- Binding of RBMX to NORAD was found to be dispensable for its genome maintenance function.
Conclusions:
- NORAD's primary role in genome maintenance is mediated through its interaction with PUMILIO proteins in the cytoplasm.
- The interaction with RBMX in the nucleus does not appear to contribute significantly to NORAD's genome stabilizing function.
- These findings highlight the NORAD-PUMILIO axis as a key player in genome maintenance.
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