Combining affinity purification and mass spectrometry to define the network of the nuclear proteins interacting with the N-terminal region of FMRP - IDEX UCA JEDI Université Côte d'Azur Accéder directement au contenu
Article Dans Une Revue Frontiers in Molecular Biosciences Année : 2022

Combining affinity purification and mass spectrometry to define the network of the nuclear proteins interacting with the N-terminal region of FMRP

Résumé

Fragile X-Syndrome (FXS) represents the most common inherited form of intellectual disability and the leading monogenic cause of Autism Spectrum Disorders. In most cases, this disease results from the absence of expression of the protein FMRP encoded by the FMR1 gene (Fragile X messenger ribonucleoprotein 1). FMRP is mainly defined as a cytoplasmic RNA-binding protein regulating the local translation of thousands of target mRNAs. Interestingly, FMRP is also able to shuttle between the nucleus and the cytoplasm. However, to date, its roles in the nucleus of mammalian neurons are just emerging. To broaden our insight into the contribution of nuclear FMRP in mammalian neuronal physiology, we identified here a nuclear interactome of the protein by combining subcellular fractionation of rat forebrains with pull‐ down affinity purification and mass spectrometry analysis. By this approach, we listed 55 candidate nuclear partners. This interactome includes known nuclear FMRP-binding proteins as Adar or Rbm14 as well as several novel candidates, notably Ddx41, Poldip3, or Hnrnpa3 that we further validated by target‐specific approaches. Through our approach, we identified factors involved in different steps of mRNA biogenesis, as transcription, splicing, editing or nuclear export, revealing a potential central regulatory function of FMRP in the biogenesis of its target mRNAs. Therefore, our work considerably enlarges the nuclear proteins interaction network of FMRP in mammalian neurons and lays the basis for exciting future mechanistic studies deepening the roles of nuclear FMRP in neuronal physiology and the etiology of the FXS.
Fichier principal
Vignette du fichier
fmolb-09-954087.pdf (2.27 Mo) Télécharger le fichier
Origine : Publication financée par une institution

Dates et versions

inserm-03836622 , version 1 (02-11-2022)

Identifiants

Citer

Félicie Kieffer, Fahd Hilal, Anne-Sophie Gay, Delphine Debayle, Marie Pronot, et al.. Combining affinity purification and mass spectrometry to define the network of the nuclear proteins interacting with the N-terminal region of FMRP. Frontiers in Molecular Biosciences, 2022, 9, pp.954087. ⟨10.3389/fmolb.2022.954087⟩. ⟨inserm-03836622⟩
54 Consultations
64 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More