# Dynamic RNA binding and unfolding by nonsense-mediated mRNA decay factor UPF2

## TL;DR

This paper explores how UPF2, a key protein in RNA decay, binds and unfolds RNA structures, potentially aiding in gene regulation.

## Contribution

The study identifies specific RNA-binding domains in UPF2 and demonstrates its RNA annealing and unfolding capabilities.

## Key findings

- UPF2's first and third MIF4G domains are main RNA/DNA-binding modules.
- UPF2 unfolds reporter hairpin RNA structures and stabilizes single-stranded RNA.
- UPF2 undergoes conformational changes upon RNA binding, suggesting functional remodeling.

## Abstract

Nonsense-mediated mRNA decay (NMD) is an mRNA surveillance pathway involved in translational control and gene expression regulation. Core NMD factors up-frameshift proteins UPF1, UPF2, and UPF3B are conserved from yeast to humans and essential to target mRNAs with a premature stop codon for decay. UPF2 binding to UPF1 activates UPF1's ATPase and helicase activities, and UPF2 binding to UPF3B is important for its association with the exon junction complex and efficient NMD. However, UPF2's association with RNA remains largely uncharacterized. Here, we analyze nucleic acid binding, identifying the first and third MIF4G domains of UPF2 as main RNA-/DNA-binding modules. We find that UPF2's MIF4G domain-3 has RNA annealing activity, while full-length UPF2 unfolds our reporter hairpin RNA structure. We show that UPF2 preferentially binds and stabilizes single-stranded RNA (ss-RNA) in a sequence-independent manner. Concomitant to ss-RNA binding, UPF2 undergoes a distinct conformational change in its otherwise highly dynamic structure. UPF2's RNA binding and unfolding activity may support UPF1's helicase and messenger ribonucleoprotein remodeling activity and, in combination with UPF3B, stabilize UPF1's association with nonsense mRNA.