HIV-1 REV NUCLEAR EXPORT SIGNAL BINDING PEPTIDES ISOLATED BY PHAGE DISPLAY

The human immunodeficiency virus type 1 (HIV-1) Rev protein is absolut ely essential in the viral replication cycle, where it induces the pro duction of viral structural proteins. Rev functions in part by inducin g the nuclear export of incompletely spliced mRNA species specified by the presence of an RNA element, the Rev response element (RRE). Sever al proteins implicated in RNA processing and nucleo-cytoplasmic transp ort have been shown to interact with Rev, however, their exact roles r emain unknown. To map potential protein recognition sites within the R ev structure, we have screened a phage library, displaying random 15-m er peptides, and isolated clones exhibiting similar sequences that spe cifically interact with Rev. The binding sites on Rev of the correspon ding synthetic peptides were characterised by protein footprinting, in volving partial proteolysis of radioactively end-labelled Rev protein. Two of the peptides produced a significant footprint within the nucle ar export signal of Rev, raising the possibility that they mimic the b inding of cellular protein factors implicated in nuclear export. (C) 1 998 Academic Press.