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Nature. 2018 Sep;561(7721):132-136. doi: 10.1038/s41586-018-0453-z. Epub 2018 Aug 27.

The NORAD lncRNA assembles a topoisomerase complex critical for genome stability.

Author information

1
Broad Institute of MIT and Harvard, Cambridge, MA, USA. mathias@broadinstitute.org.
2
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
3
Division of Hematology/Oncology, Boston Children's Hospital and Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA.
4
Program in Biological and Biomedical Sciences, Harvard University, Cambridge, MA, USA.
5
Division of Health Sciences and Technology, MIT, Cambridge, MA, USA.
6
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA, USA.
7
Broad Institute of MIT and Harvard, Cambridge, MA, USA. eric@broadinstitute.org.
8
Department of Biology, MIT, Cambridge, MA, USA. eric@broadinstitute.org.
9
Department of Systems Biology, Harvard Medical School, Boston, MA, USA. eric@broadinstitute.org.

Abstract

The human genome contains thousands of long non-coding RNAs1, but specific biological functions and biochemical mechanisms have been discovered for only about a dozen2-7. A specific long non-coding RNA-non-coding RNA activated by DNA damage (NORAD)-has recently been shown to be required for maintaining genomic stability8, but its molecular mechanism is unknown. Here we combine RNA antisense purification and quantitative mass spectrometry to identify proteins that directly interact with NORAD in living cells. We show that NORAD interacts with proteins involved in DNA replication and repair in steady-state cells and localizes to the nucleus upon stimulation with replication stress or DNA damage. In particular, NORAD interacts with RBMX, a component of the DNA-damage response, and contains the strongest RBMX-binding site in the transcriptome. We demonstrate that NORAD controls the ability of RBMX to assemble a ribonucleoprotein complex-which we term NORAD-activated ribonucleoprotein complex 1 (NARC1)-that contains the known suppressors of genomic instability topoisomerase I (TOP1), ALYREF and the PRPF19-CDC5L complex. Cells depleted for NORAD or RBMX display an increased frequency of chromosome segregation defects, reduced replication-fork velocity and altered cell-cycle progression-which represent phenotypes that are mechanistically linked to TOP1 and PRPF19-CDC5L function. Expression of NORAD in trans can rescue defects caused by NORAD depletion, but rescue is significantly impaired when the RBMX-binding site in NORAD is deleted. Our results demonstrate that the interaction between NORAD and RBMX is important for NORAD function, and that NORAD is required for the assembly of the previously unknown topoisomerase complex NARC1, which contributes to maintaining genomic stability. In addition, we uncover a previously unknown function for long non-coding RNAs in modulating the ability of an RNA-binding protein to assemble a higher-order ribonucleoprotein complex.

PMID:
30150775
DOI:
10.1038/s41586-018-0453-z
[Indexed for MEDLINE]

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