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Nup358 binds to AGO proteins through its SUMO-interacting motifs and promotes the association of target mRNA with

Manas Ranjan Sahoo1, Swati Gaikwad1, Deepak Khuperkar1

  • 1National Centre for Cell Science, S.P. Pune University Campus, Pune, India.

EMBO Reports
|January 1, 2017
PubMed

Insights

The nucleoporin Nup358 protein is crucial for microRNA (miRNA)-guided gene silencing by helping the miRNA-induced silencing complex (miRISC) find target messenger RNAs (mRNAs). Nup358

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • MicroRNA (miRNA)-guided messenger RNA (mRNA) repression by the miRNA-induced silencing complex (miRISC) is a key post-transcriptional gene silencing mechanism.
  • The precise in vivo mechanisms by which miRISC identifies target mRNAs remain incompletely understood.
  • Nucleoporins, such as Nup358, are increasingly recognized for roles beyond nuclear transport, including in gene regulation.

Purpose of the Study:

  • To investigate the role of the nucleoporin Nup358 in miRNA-mediated gene silencing.
  • To elucidate the molecular interactions and cellular localization of Nup358 in the context of the miRNA pathway.
  • To identify novel interaction motifs for Argonaute (AGO) proteins.

Main Methods:

  • Depletion of Nup358 using RNA interference or other methods.
  • Immunofluorescence microscopy to assess localization of Nup358, processing bodies (P bodies), stress granules (SGs), and miRISC components.
  • Co-immunoprecipitation assays to detect protein-protein interactions between Nup358, AGO, and GW182.
  • Analysis of miRNA target mRNA association with miRISC.

Main Results:

  • Nup358 localizes to the nuclear pore complex and cytoplasmic annulate lamellae (AL), which associate with P bodies and SGs.
  • Nup358 depletion disrupts P bodies and impairs miRNA pathway function.
  • Nup358 directly interacts with AGO proteins via its SUMO-interacting motif (SIM).
  • Nup358 promotes the association of target mRNA with miRISC, suggesting a role in target recognition and binding.
  • SIMs are identified as a novel interaction motif for AGO and PIWI proteins.

Conclusions:

  • Nup358 plays a critical role in miRNA-mediated gene silencing by facilitating target mRNA recognition and miRISC assembly.
  • The interaction between Nup358 and AGO proteins, mediated by SIM, is essential for efficient miRNA function.
  • Annulate lamellae (AL) and potentially the nuclear envelope are identified as sites for miRISC-target mRNA coupling.
Keywords:
ArgonauteNup358annulate lamellaemiRNAnucleoporin

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