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Related Experiment Video

Updated: Apr 5, 2026

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans

Published on: May 23, 2020

AUF1 promotes let-7b loading on Argonaute 2.

Je-Hyun Yoon¹, Myung Hyun Jo², Elizabeth J F White³

¹Laboratory of Genetics, National Institute on Aging-Intramural Research Program, National Institutes of Health, Baltimore, Maryland 21224, USA;

Genes & Development

|August 9, 2015

Summary

The RNA-binding protein AUF1 promotes microRNA let-7 loading onto AGO2, enhancing gene silencing. This discovery reveals a new mechanism for regulating eukaryotic gene expression via RNA-binding proteins and microRNAs.

Keywords:

Argonaute RISC hnRNP D microRNA ribonucleoprotein complex

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Area of Science:

Molecular Biology
Gene Regulation
RNA Biology

Background:

Eukaryotic gene expression is regulated post-transcriptionally by RNA-binding proteins (RBPs) and microRNAs.
AU-rich-binding factor 1 (AUF1) is an RBP involved in RNA regulation.

Purpose of the Study:

To investigate the interaction between AUF1 and microRNA let-7b.
To elucidate the role of AUF1 in microRNA-mediated gene silencing.

Main Methods:

In vitro binding assays to determine AUF1-let-7b affinity.
Ribonucleoprotein immunoprecipitation (RIP) to analyze protein-RNA interactions.
Single-molecule-binding analyses to study complex formation.
Assays to assess microRNA loading onto Argonaute 2 (AGO2) and subsequent mRNA decay.

Main Results:

AUF1 isoform p37 binds microRNA let-7b with high affinity (Kd = ~6 nM).
AUF1 facilitates the loading of let-7b onto AGO2, a key component of the RNA-induced silencing complex (RISC).
AGO2-bound let-7b triggers target mRNA decay, leading to gene silencing.

Conclusions:

AUF1 plays a novel role in promoting microRNA biogenesis and function.
AUF1-mediated transfer of let-7b to AGO2 is a critical step in let-7-elicited gene silencing.
This mechanism highlights a new pathway for post-transcriptional gene regulation by RBPs and microRNAs.