Phosphorylation, disorder, and phase separation govern the behavior of Frequency in the fungal circadian clock

Circadian clocks are composed of molecular oscillators that pace rhythms of gene expression to the diurnal cycle. Therein, transcriptional-translational negative feedback loops (TTFLs) generate oscillating levels of transcriptional repressor proteins that regulate their own gene expression. In the filamentous fungus Neurospora crassa, the proteins Frequency (FRQ), the FRQ-interacting RNA helicase (FRH) and Casein-Kinase I (CK1) form the FFC complex that represses expression of genes activated by the White-Collar complex (WCC). A key question concerns how FRQ orchestrates molecular interactions at the core of the clock despite containing little predicted tertiary structure. We present the reconstitution and biophysical characterization of FRQ and the FFC in unphosphorylated and highly phosphorylated states. Site-specific spin labeling and pulse-dipolar ESR spectroscopy provides domain-specific structural details on the full-length, 989-residue intrinsically disordered FRQ and the FFC. FR..., , , # Phosphorylation, disorder, and phase separation govern the behavior of Frequency in the fungal circadian clock

Description of the data and file structure

These files represent the raw data from the ESR, SAXS, UV-Vis, MALS, and microscopy experiments. The main folder titled \"Raw Data\" contains various sub-folders that are labeled by experiment type.

1. The \"Microscopy Images\" sub-folder contains all the microscopy images showing FRQ droplets +/- binding partners under different conditions. Within this folder, there are 3 sub-folders.

1. \"Co-LLPS\" contains co-phase-separation microscopy images with FRQ and either FRH or CK1. This folder also has the images from the CheY control.
2. \"np-FRQ and p-FRQ Concentration Titration +_- hexanediol\" contains microscopy images of FRQ at different concentrations +/- hexanediol, which is used to dissolve droplets and show that they are indeed liquid-like.
3. \"np-FRQ Cysless Temperature\" contains microscopy images of FRQ undergoi...