Redox-Responsive Zinc Finger Fidelity Switch in Homing Endonuclease and Intron Promiscuity in Oxidative Stress

It is well understood how mobile introns home to allelic sites, but how they are stimulated to transpose to ectopic locations on an evolutionary timescale is unclear [1]. Here we show that a group I intron can move to degenerate sites under oxidizing conditions. The phage T4 td intron endonuclease, I-Tevl, is responsible for this infidelity. We demonstrate that I-Tevl, which promotes mobility and is subject to autorepression [2] and translational control [3], is also regulated posttranslationally by a redox mechanism. Redox regulation is exercised by a zinc finger (ZF) in a linker that connects the catalytic domain of I-Tevl to the DNA binding domain. Four cysteines coordinate Zn2+ in the ZF, which ensures that I-Tevl cleaves its DNA substrate at a fixed distance, 23-25 nucleotides upstream of the intron insertion site [4]. We show that the fidelity of I-Tevl cleavage is controlled by redox-responsive Zn2+ cycling. When the ZF is mutated, or after exposure of the wild-type I-Tevl to H2O2, intron homing to degenerate sites is increased, likely because of indiscriminate DNA cleavage. These results suggest a mechanism for rapid intron dispersal, joining recent descriptions of the activation of biomolecular processes by oxidative stress through cysteine chemistry [5, 6].