Mapping and characterization of G-quadruplexes in monkeypox genomes

Area of Science:

• Virology
• Genomics
• Drug Discovery

Background:

• Monkeypox virus (MPXV), a double-stranded DNA virus, causes outbreaks with limited treatment options.
• Recent MPXV reemergence and global spread highlight the urgent need for effective antiviral therapies.
• G-quadruplexes are emerging targets for antiviral drug development.

Purpose of the Study:

• To identify and characterize G-quadruplex forming sequences within the MPXV genome.
• To investigate the potential of targeting MPXV G-quadruplexes for therapeutic intervention.
• To evaluate the antiviral activity of a G-quadruplex binding molecule against MPXV.

Main Methods:

• Genomic-scale mapping of MPXV isolates to identify conserved G-quadruplex forming sequences.
• Circular dichroism spectroscopy and solution small-angle X-ray scattering to assess G-quadruplex formation.
• Biochemical assays and cell biology experiments to evaluate G4-ligand interactions and antiviral efficacy.

Main Results:

• Two conserved, MPXV-exclusive putative G-quadruplex forming sequences were identified across 590 isolates.
• MPXV G-quadruplexes were shown to bind Thioflavin T and DHX36.
• The antiviral compound TMPyP4 demonstrated nanomolar affinity for MPXV G-quadruplexes and significantly reduced MPXV gene expression.

Conclusions:

• Conserved G-quadruplexes in the MPXV genome represent a promising target for novel antiviral drug development.
• The G-quadruplex binding molecule TMPyP4 shows significant potential for therapeutic application against MPXV.
• Further research into MPXV G-quadruplexes could lead to cost-effective treatments for monkeypox virus infections.