Abstract
White spot disease (WSD) is one of the most devastating viral infections of crustaceans caused by the white spot syndrome virus (WSSV). A conserved sequence WSSV131 in the DNA genome of WSSV was found to fold into a polymorphic G‐quadruplex structure. Supported by two mutant sequences with single G→T substitutions in the third G4 tract of WSSV131, circular dichroism and NMR spectroscopic analyses demonstrate folding of the wild‐type sequence into a three‐tetrad parallel topology comprising three propeller loops with a major 1 : 3 : 1 and a minor 1 : 2 : 2 loop length arrangement. A thermodynamic analysis of quadruplex formation by differential scanning calorimetry (DSC) indicates a thermodynamically more stable 1 : 3 : 1 loop isomer. DSC also revealed the formation of additional highly stable multimeric species with populations depending on potassium ion concentration.
Highlights
White spot disease (WSD) is one of the most devastating viral infections of crustaceans caused by the white spot syndrome virus (WSSV)
Screening the genome of the white spot syndrome virus, we found a conserved sequence with putative G4-forming ability termed WSSV131 (Figure 1)
Imino proton spectral regions for both mutants indicate a single quadruplex comprising twelve imino resonances with their superposition closely matching native WSSV131 imino signals with the G13T mutant in excess (Figure 2B). These results suggest the presence of a major 1:3:1 and a minor 1:2:2 loop isomer for polymorphic WSSV131
Summary
White spot disease (WSD) is one of the most devastating viral infections of crustaceans caused by the white spot syndrome virus (WSSV). The imino proton spectral region of a 1D NMR spectrum for WSSV131 reveals a set of 12 major imino resonances between 10.512.0 ppm characteristic for Hoogsteen hydrogen-bonded guanines arranged in a G-tetrad (Figure 2B). Imino proton spectral regions for both mutants indicate a single quadruplex comprising twelve imino resonances with their superposition closely matching native WSSV131 imino signals with the G13T mutant in excess (Figure 2B).
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