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Rhythmic activation and repression of the  frequency  ( frq ) gene are essential for normal function of the  Neurospora  circadian clock. WHITE COLLAR (WC) complex, the positive element of the  Neurospora  circadian system, is responsible for stimulation of  frq  transcription. We report that a C2H2 finger domain-containing protein IEC-1 and its associated chromatin remodeling complex INO80 play important roles in normal  Neurospora  circadian clock function. In  iec-1   KO   strains, circadian rhythms are abolished, and the  frq  transcript levels are increased compared to that of the wild-type strain. Similar results are observed in mutant strains of the INO80 subunits. Furthermore, ChIP data show that recruitment of the INO80 complex to the  frq  promoter is IEC-1-dependent. WC-mediated transcription of  frq  contributes to the rhythmic binding of the INO80 complex at the  frq  promoter. As demonstrated by ChIP analysis, the INO80 complex is required for the re-establishment of the dense chromatin environment at the  frq  promoter. In addition, WC-independent  frq  transcription is present in  ino80  mutants. Altogether, our data indicate that the INO80 complex suppresses  frq  transcription by re-assembling the suppressive mechanisms at the  frq  promoter after transcription of  frq .

Transcriptional repression of frequency by the IEC-1-INO80 complex is required for normal Neurospora circadian clock function