The Rev Responsive Element (RRE), a part of unspliced HIV RNA is crucial for production of infectious HIV virions. The viral protein Rev binds to RRE and facilitates transport to the cytoplasm. Intervention of this interaction disrupts the viral life cycle.
Using phage display techniques zinc finger proteins were designed to bind specifically to RRE IIB at the high affinity Rev binding site. Mutant proteins were produced and their affinities for the RNA were determined by gel shift assays and Surface Plasmon Resonance. Imino proton spectrum of RREIIB - zinc finger protein complexes showed appearances of new peaks as compared to that of the free RNA indicative of the formation of new base pairs on protein binding. The RRE-zinc finger protein binding was dependent on the protein structure since removal of Zn2+ by EDTA destroyed the protein structure (ββα motif) and disrupted the protein RNA complex. NMR solution structures of two of these zinc finger proteins have been determined and the structure of the RNA – protein complex is being solved to understand their mode of binding and delineate regions for further refinement by phage display.
Back to Frontiers in Chemistry and Medicine V (General Session)
Back to The 56th Southeast Regional Meeting 2004 (November 10-13, 2004)