Cargando…

Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency

Fragile X‐associated premature ovarian insufficiency (FXPOI) is among a family of disorders caused by expansion of a CGG trinucleotide repeat sequence located in the 5′ untranslated region (UTR) of the fragile X messenger ribonucleoprotein 1 (FMR1) gene on the X chromosome. Women with FXPOI have a d...

Descripción completa

Detalles Bibliográficos
Autores principales: Rosario, Roseanne, Stewart, Hazel L., Choudhury, Nila Roy, Michlewski, Gracjan, Charlet‐Berguerand, Nicholas, Anderson, Richard A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9828574/
https://www.ncbi.nlm.nih.gov/pubmed/36250920
http://dx.doi.org/10.1096/fj.202200468RR
_version_ 1784867304355397632
author Rosario, Roseanne
Stewart, Hazel L.
Choudhury, Nila Roy
Michlewski, Gracjan
Charlet‐Berguerand, Nicholas
Anderson, Richard A.
author_facet Rosario, Roseanne
Stewart, Hazel L.
Choudhury, Nila Roy
Michlewski, Gracjan
Charlet‐Berguerand, Nicholas
Anderson, Richard A.
author_sort Rosario, Roseanne
collection PubMed
description Fragile X‐associated premature ovarian insufficiency (FXPOI) is among a family of disorders caused by expansion of a CGG trinucleotide repeat sequence located in the 5′ untranslated region (UTR) of the fragile X messenger ribonucleoprotein 1 (FMR1) gene on the X chromosome. Women with FXPOI have a depleted ovarian follicle population, resulting in amenorrhea, hypoestrogenism, and loss of fertility before the age of 40. FXPOI is caused by expansions of the CGG sequence to lengths between 55 and 200 repeats, known as a FMRI premutation, however the mechanism by which the premutation drives disease pathogenesis remains unclear. Two main hypotheses exist, which describe an mRNA toxic gain‐of‐function mechanism or a protein‐based mechanism, where repeat‐associated non‐AUG (RAN) translation results in the production of an abnormal protein, called FMRpolyG. Here, we have developed an in vitro granulosa cell model of the FMR1 premutation by ectopically expressing CGG‐repeat RNA and FMRpolyG protein. We show that expanded CGG‐repeat RNA accumulated in intranuclear RNA structures, and these aggregates were able to cause significant granulosa cell death independent of FMRpolyG expression. Using an innovative RNA pulldown, mass spectrometry‐based approach we have identified proteins that are specifically sequestered by CGG RNA aggregates in granulosa cells in vitro, and thus may be deregulated as consequence of this interaction. Furthermore, we have demonstrated reduced expression of three proteins identified via our RNA pulldown (FUS, PA2G4 and TRA2β) in ovarian follicles in a FMR1 premutation mouse model. Collectively, these data provide evidence for the contribution of an mRNA gain‐of‐function mechanism to FXPOI disease biology.
format Online
Article
Text
id pubmed-9828574
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher John Wiley and Sons Inc.
record_format MEDLINE/PubMed
spelling pubmed-98285742023-01-10 Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency Rosario, Roseanne Stewart, Hazel L. Choudhury, Nila Roy Michlewski, Gracjan Charlet‐Berguerand, Nicholas Anderson, Richard A. FASEB J Research Articles Fragile X‐associated premature ovarian insufficiency (FXPOI) is among a family of disorders caused by expansion of a CGG trinucleotide repeat sequence located in the 5′ untranslated region (UTR) of the fragile X messenger ribonucleoprotein 1 (FMR1) gene on the X chromosome. Women with FXPOI have a depleted ovarian follicle population, resulting in amenorrhea, hypoestrogenism, and loss of fertility before the age of 40. FXPOI is caused by expansions of the CGG sequence to lengths between 55 and 200 repeats, known as a FMRI premutation, however the mechanism by which the premutation drives disease pathogenesis remains unclear. Two main hypotheses exist, which describe an mRNA toxic gain‐of‐function mechanism or a protein‐based mechanism, where repeat‐associated non‐AUG (RAN) translation results in the production of an abnormal protein, called FMRpolyG. Here, we have developed an in vitro granulosa cell model of the FMR1 premutation by ectopically expressing CGG‐repeat RNA and FMRpolyG protein. We show that expanded CGG‐repeat RNA accumulated in intranuclear RNA structures, and these aggregates were able to cause significant granulosa cell death independent of FMRpolyG expression. Using an innovative RNA pulldown, mass spectrometry‐based approach we have identified proteins that are specifically sequestered by CGG RNA aggregates in granulosa cells in vitro, and thus may be deregulated as consequence of this interaction. Furthermore, we have demonstrated reduced expression of three proteins identified via our RNA pulldown (FUS, PA2G4 and TRA2β) in ovarian follicles in a FMR1 premutation mouse model. Collectively, these data provide evidence for the contribution of an mRNA gain‐of‐function mechanism to FXPOI disease biology. John Wiley and Sons Inc. 2022-10-17 2022-11 /pmc/articles/PMC9828574/ /pubmed/36250920 http://dx.doi.org/10.1096/fj.202200468RR Text en © 2022 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Rosario, Roseanne
Stewart, Hazel L.
Choudhury, Nila Roy
Michlewski, Gracjan
Charlet‐Berguerand, Nicholas
Anderson, Richard A.
Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency
title Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency
title_full Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency
title_fullStr Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency
title_full_unstemmed Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency
title_short Evidence for a fragile X messenger ribonucleoprotein 1 (FMR1) mRNA gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile X‐associated premature ovarian insufficiency
title_sort evidence for a fragile x messenger ribonucleoprotein 1 (fmr1) mrna gain‐of‐function toxicity mechanism contributing to the pathogenesis of fragile x‐associated premature ovarian insufficiency
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9828574/
https://www.ncbi.nlm.nih.gov/pubmed/36250920
http://dx.doi.org/10.1096/fj.202200468RR
work_keys_str_mv AT rosarioroseanne evidenceforafragilexmessengerribonucleoprotein1fmr1mrnagainoffunctiontoxicitymechanismcontributingtothepathogenesisoffragilexassociatedprematureovarianinsufficiency
AT stewarthazell evidenceforafragilexmessengerribonucleoprotein1fmr1mrnagainoffunctiontoxicitymechanismcontributingtothepathogenesisoffragilexassociatedprematureovarianinsufficiency
AT choudhurynilaroy evidenceforafragilexmessengerribonucleoprotein1fmr1mrnagainoffunctiontoxicitymechanismcontributingtothepathogenesisoffragilexassociatedprematureovarianinsufficiency
AT michlewskigracjan evidenceforafragilexmessengerribonucleoprotein1fmr1mrnagainoffunctiontoxicitymechanismcontributingtothepathogenesisoffragilexassociatedprematureovarianinsufficiency
AT charletberguerandnicholas evidenceforafragilexmessengerribonucleoprotein1fmr1mrnagainoffunctiontoxicitymechanismcontributingtothepathogenesisoffragilexassociatedprematureovarianinsufficiency
AT andersonricharda evidenceforafragilexmessengerribonucleoprotein1fmr1mrnagainoffunctiontoxicitymechanismcontributingtothepathogenesisoffragilexassociatedprematureovarianinsufficiency