Targeting the Tat-binding site of bovine immunodeficiency virus TAR RNA with a shape-selective rhodium complex.
The Tat-binding site of the bovine immunodeficiency virus TAR RNA hairpin has been targeted by Rh(phen)2phi3+ (phen = phenanthroline, phi = 9,10-phenanthrenequinone diimine), a photochemical probe of RNA tertiary structure. The primary site cleaved by the rhodium complex, upon photoactivation, is U24, a base which participates in the novel base triple (with bases A13 and U10) characteristic of this folded RNA. delta-Rh(phen)2phi3+ binds to this site with an affinity of 2 x 10(6) M-1. Upon mutation of U24 and A13 to A24 and U13, respectively, so that the RNA oligomer is unable to form the base triple, site-specific cleavage by the rhodium complex is abolished. Moreover, as determined through rhodium photocleavage, at a concentration of 20 microM, Rh(phen)2phi3+ inhibits specific binding of BIV-Tat peptide (2 microM) to its target site. Thus the rhodium complex, in matching its shape to the opened major groove of the properly folded RNA, specifically targets its site and is able to compete for its target with the BIV-Tat peptide.
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Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125, USA.
Barton
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JK
D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism*; D001665:Binding Sites; D002417:Cattle; D015696:Gene Products, tat / Q000378:metabolism*; D016087:Immunodeficiency Virus, Bovine / Q000378:metabolism*; D007700:Kinetics; D009690:Nucleic Acid Conformation; D009942:Organometallic Compounds / Q000378:metabolism* / Q000494:pharmacology; D010616:Phenanthrenes / Q000378:metabolism* / Q000494:pharmacology; D010618:Phenanthrolines / Q000378:metabolism* / Q000494:pharmacology; D012367:RNA, Viral / Q000378:metabolism*; D012238:Rhodium / Q000378:metabolism* / Q000494:pharmacology; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D014764:Viral Proteins / Q000378:metabolism*
D000998:Antiviral Agents; D015696:Gene Products, tat; D009942:Organometallic Compounds; D010616:Phenanthrenes; D010618:Phenanthrolines; D012367:RNA, Viral; D014764:Viral Proteins; C073959:bis(phenanthroline)(phenanthrenequinone diimine)rhodium(III); D012238:Rhodium