PROBING THE STRUCTURE OF HIV-1 REV BY PROTEIN FOOTPRINTING OF MULTIPLE MONOCLONAL ANTIBODY-BINDING SITES

Human immunodeficiency virus type 1 (HIV-1) Rev is a small RNA-binding protein which is essential far viral replication. To investigate the structure of Rev we have mapped the binding sites of a panel of monocl onal antibodies (mAb) by protein footprinting and identified a mAb pro tecting amino acids within both the N- and C-terminal parts of Rev. Ou r mapping results support a previously proposed structure (Auer et al. , Biochemistry, 33 (1994) 2988-2996) predicting that a helix-loop-heli x motif in Rev brings the termini of the protein into proximity. Furth ermore, we demonstrate that the binding sites mapped by protein footpr inting are in agreement with conventional epitope mapping results and that this technique are provides an advantageous strategy for mapping discontinuous sites. (C) 1997 Federation of European Biochemical Socie ties.