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Inhibition of Moloney murine leukemia virus integration using polyamides targeting the long-terminal repeat sequences
Fan Yang1, Jason M Belitsky, Rodrigo A Villanueva
1Department of Biochemistry, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway 08854, USA.
Abstract:
The retroviral integrase (IN) carries out the integration of the viral DNA into the host genome. Both IN and the DNA sequences at the viral long-terminal repeat (LTR) are required for the integration function. In this report, a series of minor groove binding hairpin polyamides targeting sequences within terminal inverted repeats of the Moloney murine leukemia virus (M-MuLV) LTR were synthesized, and their effects on integration were analyzed. Using cell-free in vitro integration assays, polyamides targeting the conserved CA dinucleotide with cognate sites closest to the terminal base pairs were effective at blocking 3' processing but not strand transfer. Polyamides which efficiently inhibited 3' processing and strand transfer targeted the LTR sequences through position 9. Polyamides that inhibited integration were effective at nanomolar concentrations and showed subnanomolar affinity for their cognate LTR sites. These studies highlight the role of minor groove interactions within the LTR termini for retroviral integration.
Insights
Researchers developed novel polyamides to block retroviral DNA integration. These molecules target Moloney murine leukemia virus (M-MuLV) long-terminal repeat (LTR) sequences, effectively inhibiting key integration steps.
Area of Science:
- Molecular Biology
- Virology
- Drug Discovery
Background:
- Retroviral DNA integration into the host genome is facilitated by the retroviral integrase (IN) enzyme.
- Both the integrase enzyme and specific DNA sequences within the viral long-terminal repeat (LTR) are essential for this integration process.
Purpose of the Study:
- To synthesize and evaluate minor groove binding hairpin polyamides targeting Moloney murine leukemia virus (M-MuLV) LTR sequences.
- To determine the effects of these polyamides on the retroviral integration process, specifically focusing on 3' processing and strand transfer.
Main Methods:
- Synthesis of a series of minor groove binding hairpin polyamides.
- Cell-free in vitro integration assays to analyze the effects of polyamides on M-MuLV integration.
- Affinity measurements of polyamides to their cognate LTR sites.
Main Results:
- Polyamides targeting the conserved CA dinucleotide near terminal base pairs blocked 3' processing but not strand transfer.
- Polyamides targeting LTR sequences up to position 9 inhibited both 3' processing and strand transfer.
- Effective integration inhibition was observed at nanomolar concentrations with subnanomolar affinity.
Conclusions:
- Minor groove interactions within LTR termini play a critical role in retroviral integration.
- Developed polyamides demonstrate potential as inhibitors of retroviral integration by targeting specific LTR sequences.