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Codon Optimization> | The plasmid pHyPerRedSc was constructed by synthesizing the HyPerRed H2O2 sensor33, codon optimized for yeast (sequence available upon request) and subcloning it into the vector pRS416-GPD62 using BamHI and EcoRI sites inserted upstream and downstream of the HyPerRedSc open reading frame, respectively (GenScript).... C199S point mutagenesis was performed by changing a TGT codon into a TCT in the HyPerRedSc coding sequence (positions 361–363, GenScript). | Get A Quote |
Yeast lacks dedicated photoreceptors; however, blue light still causes pronounced oscillations of the transcription factor Msn2 into and out of the nucleus. Here we show that this poorly understood phenomenon is initiated by a peroxisomal oxidase, which converts light into a hydrogen peroxide (H2O2) signal that is sensed by the peroxiredoxin Tsa1 and transduced to thioredoxin, to counteract PKA-dependent Msn2 phosphorylation. Upon H2O2, the nuclear retention of PKA catalytic subunits, which contributes to delayed Msn2 nuclear concentration, is antagonized in a Tsa1-dependent manner. Conversely, peroxiredoxin hyperoxidation interrupts the H2O2 signal and drives Msn2 oscillations by superimposing on PKA feedback ... More