DNA-binding and antiviral activity of bis-netropsins containing clusters of lysine residues in the N-terminal region

Андронова, В. Л.; Гроховский, С. Л.; Surovaya, A. N.; Gurskii, G. V.; Галегов, Г. А.

doi:10.1007/s10628-005-0016-8

Cited by 2 publications

(1 citation statement)

References 14 publications

(20 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the present paper, we provide experimental evidence that complexes of the herpes simplex virus type 1 (HSV1) OBP with the viral replication origins, two OriS sites and one OriL site, serve as perspective targets for suppression of the diseases caused by HSV1. Previous studies showed that bis-linked netropsin derivatives effectively suppress development of virus-induced cytopathic effect in the cultured cells ( Andronova et al, 2008 ; Andronova, Grokhovsky, Surovaya, Gursky, & Galegov, 2001 , 2004 ; Lown et al, 1989 ). They also exert therapeutic effects on laboratory animals infected by HSV1 ( Andronova et al, 2012 ; Andronova, Grokhovsky, Surovaya, Gursky, & Galegov, 2007 ).…”

Section: Introductionmentioning

confidence: 99%

Complex of the herpes simplex virus type 1 origin binding protein UL9 with DNA as a platform for the design of a new type of antiviral drugs

Bazhulina

Surovaya

Gursky

et al. 2013

Journal of Biomolecular Structure and Dynamics

View full text Add to dashboard Cite

The herpes simplex virus type 1 origin-binding protein, OBP, is a DNA helicase encoded by the UL9 gene. The protein binds in a sequence-specific manner to the viral origins of replication, two OriS sites and one OriL site. In order to search for efficient inhibitors of the OBP activity, we have obtained a recombinant origin-binding protein expressed in Escherichia coli cells. The UL9 gene has been amplified by PCR and inserted into a modified plasmid pET14 between NdeI and KpnI sites. The recombinant protein binds to Box I and Box II sequences and possesses helicase and ATPase activities. In the presence of ATP and viral protein ICP8 (single-strand DNA-binding protein), the initiator protein induces unwinding of the minimal OriS duplex (≈80 bp). The protein also binds to a single-stranded DNA (OriS*) containing a stable Box I-Box III hairpin and an unstable AT-rich hairpin at the 3′-end. In the present work, new minor groove binding ligands have been synthesized which are capable to inhibit the development of virus-induced cytopathic effect in cultured Vero cells. Studies on binding of these compounds to DNA and synthetic oligonucleotides have been performed by fluorescence methods, gel mobility shift analysis and footprinting assays. Footprinting studies have revealed that Pt-bis-netropsin and related molecules exhibit preferences for binding to the AT-spacer in OriS. The drugs stabilize structure of the AT-rich region and inhibit the fluctuation opening of AT-base pairs which is a prerequisite to unwinding of DNA by OBP. Kinetics of ATP-dependent unwinding of OriS in the presence and absence of netropsin derivatives have been studied by measuring the efficiency of Forster resonance energy transfer (FRET) between fluorophores attached to 5′- and 3′- ends of an oligonucleotide in the minimal OriS duplex. The results are consistent with the suggestion that OBP is the DNA Holiday junction (HJ) binding helicase. The protein induces conformation changes (bending and partial melting) of OriS duplexes and stimulates HJ formation in the absence of ATP. The antiviral activity of bis-netropsins is coupled with their ability to inhibit the fluctuation opening of AT base pairs in the A + T cluster and their capacity to stabilize the structure of the AT-rich hairpin in the single-stranded oligonucleotide corresponding to the upper chain in the minimal duplex OriS. The antiviral activities of bis-netropsins in cell culture and their therapeutic effects on HSV1-infected laboratory animals have been studied.

show abstract