The miRISC component AGO2 has multiple binding sites for Nup358 SUMO-interacting motif

Prachi Deshmukh; Shubha Markande; Vikas Fandade; Yogendra Ramtirtha; Mallur Srivatsan Madhusudhan; Jomon Joseph

doi:10.1016/j.bbrc.2021.03.140

The miRISC component AGO2 has multiple binding sites for Nup358 SUMO-interacting motif

Biochem Biophys Res Commun. 2021 Jun 4:556:45-52. doi: 10.1016/j.bbrc.2021.03.140. Epub 2021 Apr 7.

Authors

Prachi Deshmukh¹, Shubha Markande¹, Vikas Fandade¹, Yogendra Ramtirtha², Mallur Srivatsan Madhusudhan², Jomon Joseph³

Affiliations

¹ National Centre for Cell Science, S.P. Pune University Campus, Pune, India.
² Division of Biology, Indian Institute of Science Education and Research, Pune, India.
³ National Centre for Cell Science, S.P. Pune University Campus, Pune, India. Electronic address: josephj@nccs.res.in.

PMID: 33838501
DOI: 10.1016/j.bbrc.2021.03.140

Abstract

Micro-RNA mediated suppression of mRNA translation represents a major regulatory mode of post-transcriptional gene expression. Recently, the nucleoporin Nup358 was shown to interact with AGO protein, a key component of miRNA-induced silencing complex (miRISC), and facilitate the coupling of miRISC with target mRNA. Previous results suggested that SUMO-interacting motifs (SIMs) present on Nup358 mediate interaction with AGO protein. Here we show that Nup358-SIM has multiple interacting regions on AGO2, specifically within the N, PAZ and MID domains, with an affinity comparable to SIM-SUMO1 interaction. The study also unraveled specific residues involved in the interaction of AGO2 with miRNA-loading components such as Dicer and HSP90. Collectively, the results support the conclusion that multiple SIMs contribute to the association of Nup358 with AGO2.

Keywords: AGO2; Dicer; Hsp90; Nup358; SUMO-interaction motif; miRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Motifs
Argonaute Proteins / chemistry*
Argonaute Proteins / genetics
Argonaute Proteins / metabolism*
Binding Sites
DEAD-box RNA Helicases / metabolism
HEK293 Cells
HSP90 Heat-Shock Proteins / metabolism
Humans
Molecular Chaperones / chemistry*
Molecular Chaperones / metabolism*
Nuclear Pore Complex Proteins / chemistry*
Nuclear Pore Complex Proteins / metabolism*
Peptide Fragments / chemistry
Peptide Fragments / genetics
Peptide Fragments / metabolism
Point Mutation
Protein Binding
Protein Domains
Ribonuclease III / metabolism
SUMO-1 Protein / metabolism*
Sequence Deletion
Surface Plasmon Resonance

Substances

AGO2 protein, human
Argonaute Proteins
HSP90 Heat-Shock Proteins
Molecular Chaperones
Nuclear Pore Complex Proteins
Peptide Fragments
SUMO-1 Protein
ran-binding protein 2
DICER1 protein, human
Ribonuclease III
DEAD-box RNA Helicases