PUMILIO, but not RBMX, binding is required for regulation of genomic stability by noncoding RNA NORAD

Mahmoud M. Elguindy, Florian Kopp, Mohammad Goodarzi, Frederick Rehfeld, Anu Thomas, Tsung-Cheng Chang, Joshua T Mendell

Research output: Contribution to journalArticle

Abstract

NORAD is a conserved long noncoding RNA (lncRNA) that is required for genome stability in mammals. NORAD acts as a negative regulator of PUMILIO (PUM) proteins in the cytoplasm, and we previously showed that loss of NORAD or PUM hyperactivity results in genome instability and premature aging in mice (Kopp et al., 2019). Recently, however, it was reported that NORAD regulates genome stability through an interaction with the RNA binding protein RBMX in the nucleus. Here, we addressed the contributions of NORAD:PUM and NORAD:RBMX interactions to genome maintenance by this lncRNA in human cells. Extensive RNA FISH and fractionation experiments established that NORAD localizes predominantly to the cytoplasm with or without DNA damage. Moreover, genetic rescue experiments demonstrated that PUM binding is required for maintenance of genomic stability by NORAD whereas binding of RBMX is dispensable for this function. These data provide an important foundation for further mechanistic dissection of the NORAD-PUMILIO axis in genome maintenance.

Original languageEnglish (US)
Article numbere48625
JournaleLife
Volume8
DOIs
StatePublished - Jul 1 2019

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Untranslated RNA
Genomic Instability
Genes
Long Noncoding RNA
Maintenance
Cytoplasm
Genome
Premature Aging
RNA-Binding Proteins
Dissection
Mammals
DNA Damage
Fractionation
RNA
Aging of materials
Experiments
Cells
DNA
Proteins

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

PUMILIO, but not RBMX, binding is required for regulation of genomic stability by noncoding RNA NORAD. / Elguindy, Mahmoud M.; Kopp, Florian; Goodarzi, Mohammad; Rehfeld, Frederick; Thomas, Anu; Chang, Tsung-Cheng; Mendell, Joshua T.

In: eLife, Vol. 8, e48625, 01.07.2019.

Research output: Contribution to journalArticle

Elguindy, Mahmoud M. ; Kopp, Florian ; Goodarzi, Mohammad ; Rehfeld, Frederick ; Thomas, Anu ; Chang, Tsung-Cheng ; Mendell, Joshua T. / PUMILIO, but not RBMX, binding is required for regulation of genomic stability by noncoding RNA NORAD. In: eLife. 2019 ; Vol. 8.
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abstract = "NORAD is a conserved long noncoding RNA (lncRNA) that is required for genome stability in mammals. NORAD acts as a negative regulator of PUMILIO (PUM) proteins in the cytoplasm, and we previously showed that loss of NORAD or PUM hyperactivity results in genome instability and premature aging in mice (Kopp et al., 2019). Recently, however, it was reported that NORAD regulates genome stability through an interaction with the RNA binding protein RBMX in the nucleus. Here, we addressed the contributions of NORAD:PUM and NORAD:RBMX interactions to genome maintenance by this lncRNA in human cells. Extensive RNA FISH and fractionation experiments established that NORAD localizes predominantly to the cytoplasm with or without DNA damage. Moreover, genetic rescue experiments demonstrated that PUM binding is required for maintenance of genomic stability by NORAD whereas binding of RBMX is dispensable for this function. These data provide an important foundation for further mechanistic dissection of the NORAD-PUMILIO axis in genome maintenance.",
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AU - Goodarzi, Mohammad

AU - Rehfeld, Frederick

AU - Thomas, Anu

AU - Chang, Tsung-Cheng

AU - Mendell, Joshua T

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