title: Mechanism of RNA cleavage catalyzed by sequence specific polyamide nucleic acid-neamine conjugate.
abstract: In earlier studies, we found that a conjugate of neamine-polyamide nucleic acid targeting transactivation response element of HIV-1 RNA genome (HIV-1 TAR) displayed anti-HIV-1 activity and sequence-specific cleavage of the target RNA in vitro. Here we show that both the position of conjugation of polyamide nucleic acid (PNA) on neamine and the length of the spacer are critical parameters for conferring cleavage activity to the conjugate. The conjugation of PNA via a spacer incorporating 11 atoms to the 5-position of ring I of the neamine core conferred sequence-specific RNA cleavage activity on the conjugate, while conjugation to the 4'-position of ring II abolished this activity. Similarly, 5-neamine PNA complementary to TAR sequence of HIV-1 genome (PNA(TAR)) conjugates having either a 23-atom spacer or a bulky dansyl group between PNA and the neamine core also resulted in complete loss of cleavage activity. Based on these observations, we propose a mechanism for the observed RNA cleavage catalyzed by the conjugate involving unprotonated and protonated amino groups at the 3-position of ring I and the 6'-position of ring II of the neamine core, respectively.
authors.affiliation: Department of Biochemistry and Molecular Biology, UMDNJ-New Jersey Medical School, Newark, NJ 07103, USA.
authors.forename: Binay
authors.initials: B
authors.lastname: Chaubey
authors.forename: Snehlata
authors.initials: S
authors.lastname: Tripathi
authors.forename: Jérome
authors.initials: J
authors.lastname: Désiré
authors.forename: Isabelle
authors.initials: I
authors.lastname: Baussanne
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Décout
authors.forename: Virendra N
authors.initials: VN
authors.lastname: Pandey
mesh_terms: D000617:Aminoglycosides / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D005601:Framycetin / Q000737:chemistry / Q000378:metabolism*; D016325:HIV Long Terminal Repeat / Q000235:genetics*; D015497:HIV-1 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009757:Nylons / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000737:chemistry* / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D020218:Response Elements / Q000235:genetics
chemical_list: D000617:Aminoglycosides; D009757:Nylons; D016376:Oligonucleotides, Antisense; D012367:RNA, Viral; D005601:Framycetin; C488396:neamine