mRNA decay is critical for the regulation of gene expression and the quality control of mRNA. RNA helicases play a key role in eukaryotic mRNA decay. In general, RNA helicases utilize the energy of ATP hydrolysis to remodel RNA or RNA-protein complexes, resulting in the separation of RNA duplex strand and/or displacement of proteins from the RNA molecule in RNP (ribonucleoprotein) complexes. Recently, high-resolution crystal structures of RNA helicases in mRNA decay have contributed a great deal to our understanding of these key molecules. In the present review, we focus on the structural and mechanistic aspects of three RNA helicases, Dhh1, Upf1 and eIF4AIII, that are involved in eukaryotic mRNA decay.