Hsp70 is a protein chaperone that prevents protein aggregation and aids protein folding by binding to hydrophobic peptide domains through a reversible mechanism directed by an ATPase cycle. However， Hsp70 also binds U-rich RNA including some AU-rich elements (AREs) that regulate the decay kinetics of select mRNAs and has recently been shown to bind and stabilize some ARE-containing transcripts in cells. Previous studies indicated that both the ATP- and peptide-binding domains of Hsp70 contributed to the stability of Hsp70-RNA complexes and that ATP might inhibit RNA recruitment. This suggested the possibility that RNA binding by Hsp70 might mimic features of its peptide-directed chaperone activities. Here， using purified， cofactor-free preparations of recombinant human Hsp70 and quantitative biochemical approaches， we found that high-affinity RNA binding requires at least 30 nucleotides of RNA sequence but is independent of Hsp70's nucleotide-bound status， ATPase activity， or peptide-binding roles. Furthermore， although both the ATP- and peptide-binding domains of Hsp70 could form complexes with an ARE sequence from mRNA ， only the peptide-binding domain could recover cellular mRNA in ribonucleoprotein immunoprecipitations. Finally， Hsp70-directed stabilization of mRNA in cells was mediated exclusively by the protein's peptide-binding domain. Together， these findings indicate that the RNA-binding and mRNA-stabilizing functions of Hsp70 are independent of its protein chaperone cycle but also provide potential mechanical explanations for several well-established and recently discovered cytoprotective and RNA-based Hsp70 functions.