Aminoglycoside-Arginine Conjugates That Bind TAR RNA: Synthesis, Characterization, and Antiviral Activity
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- 作者：Alexander Litovchick ; Artem G. Evdokimov ; and Aviva Lapidot
- 刊名：Biochemistry
- 出版年：2000
- 出版时间：March 21, 2000
- 年：2000
- 卷：39
- 期：11
- 页码：2838 - 2852
- 全文大小：714K
- 年卷期：v.39,no.11(March 21, 2000)
- ISSN：1520-4995
文摘
Regulation of HIV gene expression is crucially dependent on binding of the trans-activatorprotein, Tat, to the trans-activation response RNA element, TAR, found at the 5' end of all HIV-1 transcripts.Tat-TAR interaction is mediated by a short arginine-rich domain of the protein. Disruption of thisinteraction could, in theory, create a state of complete viral latency. A new class of small-moleculepeptidomimetic TAR RNA binders, conjugates of aminoglycosides and arginine, was recently designed[Litovchick, A., Evdokimov, A. G., and Lapidot, A. (1999) FEBS Lett. 445, 73-79]. Two of thesecompounds, the tri-arginine derivative of gentamicin C (R3G) and the tetra-arginine derivative of kanamycinA (R4K), bind efficiently and specifically to TAR RNA. These compounds display negligible toxicitywhile being transported and accumulated in cell nuclei. Here we present a detailed synthesis and chemicalcharacterization of the aminoglycoside-arginine conjugates R3G and R4K as well as GB4K, the tetra-
-guanidinobutyric derivative of kanamycin A. Their binding sites on TAR RNA were assigned by RNaseA, uranyl nitrate, and lead acetate footprinting. The conjugates interact with TAR RNA in the widenedmajor groove, formed by the UCU bulge and the neighboring base pairs of the upper stem portion ofTAR, the binding site of Tat protein, and Tat-derived peptides (e.g., R52). Our results suggest an additionalbinding site of R4K and R3G compounds, in the lower stem-bulge region of TAR. The antiviral activityof the conjugates in cultured equine dermal fibroblasts infected with equine infectious anemia virus, usedas a model system of HIV-infected cells, is also presented.
-guanidinobutyric derivative of kanamycin A. Their binding sites on TAR RNA were assigned by RNaseA, uranyl nitrate, and lead acetate footprinting. The conjugates interact with TAR RNA in the widenedmajor groove, formed by the UCU bulge and the neighboring base pairs of the upper stem portion ofTAR, the binding site of Tat protein, and Tat-derived peptides (e.g., R52). Our results suggest an additionalbinding site of R4K and R3G compounds, in the lower stem-bulge region of TAR. The antiviral activityof the conjugates in cultured equine dermal fibroblasts infected with equine infectious anemia virus, usedas a model system of HIV-infected cells, is also presented.