title: Aromatic polyamidines inhibiting the Tat-induced HIV-1 transcription recognize structured TAR-RNA.
abstract: We have investigated the effects of aromatic polyamidines on HIV-1 transcription. We found a block to Tat-induced HIV-1 transcription assessed by inhibition of CAT activity in HL3T1 cells at a concentration lower than the IC50 value, suggesting that molecules with three (TAPB) and four (TAPP) benzamidine rings could be useful against HIV-1. In contrast, aromatic polyamidines with only two benzamidine rings (DAPP) did not block Tat-induced transcription. We reasoned that this effect could be due to binding of TAPB and TAPP to HIV-1 TAR RNA. By EMSA and filter binding assays, we studied possible interactions of aromatic polyamidines with HIV-1 TAR RNA. Wild-type TAR RNA or TAR RNA with mutations in the stem or bulge sequences, but retaining the stem-loop structure, was used to define the RNA-binding activities of these compounds. Our data suggest that aromatic polyamidines with two (DAPP) and four (TAPP) benzamidine rings, respectively, do not bind to TAR RNA or bind without sequence selectivity. Interestingly, an aromatic polyamidine with three benzamidine rings (TAPB) recognizes the wild-type TAR RNA in a specific manner. Furthermore, we found that introduction of one halogen atom into the benzamidine rings strongly increases the RNA-binding activity of these compounds.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Ferrara, Italy. msc@unife.it
authors.forename: C
authors.initials: C
authors.lastname: Mischiati
authors.forename: K T
authors.initials: KT
authors.lastname: Jeang
authors.forename: G
authors.initials: G
authors.lastname: Feriotto
authors.forename: L
authors.initials: L
authors.lastname: Breda
authors.forename: M
authors.initials: M
authors.lastname: Borgatti
authors.forename: N
authors.initials: N
authors.lastname: Bianchi
authors.forename: R
authors.initials: R
authors.lastname: Gambari
mesh_terms: D019380:Anti-HIV Agents / Q000737:chemistry / Q000494:pharmacology*; D001550:Benzamidines / Q000494:pharmacology*; D002460:Cell Line / Q000821:virology; D015195:Drug Design; D024202:Electrophoretic Mobility Shift Assay; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D015696:Gene Products, tat / Q000037:antagonists & inhibitors*; D016325:HIV Long Terminal Repeat / Q000187:drug effects* / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D019169:Jurkat Cells / Q000821:virology; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry*; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000821:virology; D015533:Transcriptional Activation / Q000187:drug effects*; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D019380:Anti-HIV Agents; D001550:Benzamidines; D015696:Gene Products, tat; D012367:RNA, Viral; D054322:tat Gene Products, Human Immunodeficiency Virus; C074442:1,3-bis(4-amidinophenoxy)-2-(4-amidinophenoxymethyl)ethylpropane; C057885:1,3-di-(4-amidinophenoxy)-2,2-bis-(4-amidinophenoxymethyl)propane; C005555:propamidine