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RESEARCH PRODUCT

Inhibition of Formation of Rev-RRE Complex by Pyronin Y

H. UshijimaAndreas BekHeinz-christoph SchröderHelmut MerzW. E. G. MüllerKarin Pfeifer

subject

0301 basic medicineStereochemistryviruses030106 microbiologyResponse elementIntercalation (chemistry)RNAGeneral MedicineBiology01 natural sciencesMolecular biologyIn vitroVirus0104 chemical scienceslaw.inventionDissociation constant010404 medicinal & biomolecular chemistry03 medical and health sciencesMechanism of actionlawmedicineRecombinant DNAmedicine.symptom

description

The interaction of pyronin Y, an RNA intercalating drug, with the binding of Rev protein from human immunodeficiency virus type 1 (HIV-1) to Rev-responsive element (RRE)-containing env RNA was studied. In gel retardation assays, recombinant Rev protein tightly bound to in vitro transcribed RRE RNA. Nitrocellulose-filter-binding studies revealed a dissociation constant of ≈(1–2) = 10−10M (Pfeifer et al., 1991). Pyronin Y efficiently suppressed formation of the Rev-RRE complex. At a concentration of 1 μg ml−1, complex formation was almost completely inhibited. Electron microscopy showed that Rev oligomerizes in the presence of RRE-containing RNA with the formation of short rod-like structures or long filaments, depending on the length of the transcript. Assembly of Rev protein along RRE-containing RNAs was abolished after addition of pyronin Y. Thus pyronin Y represents the first compound described to inhibit Rev-RRE complex formation.

https://doi.org/10.1177/095632029300400205