Binding characteristics of small molecules that mimic nucleocapsid protein-induced maturation of stem-loop 1 of HIV-1 RNA.
As a retrovirus, the human immunodeficiency virus (HIV-1) packages two copies of the RNA genome as a dimer in the infectious virion. Dimerization is initiated at the dimer initiation site (DIS) which encompasses stem-loop 1 (SL1) in the 5'-UTR of the genome. Study of genomic dimerization has been facilitated by the discovery that short RNA fragments containing SL1 can dimerize spontaneously without any protein factors. On the basis of the palindromic nature of SL1, a kissing loop model has been proposed. First, a metastable kissing dimer is formed via standard Watson-Crick base pairs and then converted into a more stable extended dimer by the viral nucleocapsid protein (NCp7). This dimer maturation in vitro is believed to mimic initial steps in the RNA maturation in vivo, which is correlated with viral infectivity. We previously discovered a small molecule activator, Lys-Ala-7-amido-4-methylcoumarin (KA-AMC), which facilitates dimer maturation in vitro, and determined aspects of its structure-activity relationship. In this report, we present measurements of the binding affinity of the activators and characterization of their interactions with the SL1 RNA. Guanidinium groups and increasing positive charge on the side chain enhance affinity and activity, but features in the aromatic ring at least partially decouple affinity from activity. Although KA-AMC can bind to multiple structural motifs, the NMR study showed KA-AMC preferentially binds to unique structural motifs, such as the palindromic loop and the G-rich internal loop in the SL1 RNA. NCp7 binds to SL1 only 1 order of magnitude more tightly than the best small molecule ligand tested. This study provides guidelines for the design of superior small molecules that bind to the SL1 RNA that have the potential of being developed as an antiviral by interfering with SL1-NCp7 interaction at the packaging and/or maturation stages.
Chung
Janet
J
Graduate Group in Chemistry and Chemical Biology, University of California-San Francisco, 600 16th Street, San Francisco, CA 94158-2517, USA.
Ulyanov
Nikolai B
NB
Guilbert
Christophe
C
Mujeeb
Anwer
A
James
Thomas L
TL
D001665:Binding Sites; D003374:Coumarins / Q000494:pharmacology; D019281:Dimerization; D004151:Dipeptides / Q000494:pharmacology; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D018716:Molecular Mimicry; D009690:Nucleic Acid Conformation / Q000187:drug effects; D019590:Nucleocapsid Proteins / Q000494:pharmacology*; D012367:RNA, Viral / Q000737:chemistry* / Q000187:drug effects; D013329:Structure-Activity Relationship
D003374:Coumarins; D004151:Dipeptides; D019590:Nucleocapsid Proteins; D012367:RNA, Viral; C045792:7-lysylalanyl-4-methylcoumarinamide