A prion-like domain in ELF3 functions as a thermosensor inArabidopsis

The adaptability of the plantArabidopsis thalianato different temperatures is regulated by the ability of its ELF3 protein to undergo liquid-liquid phase separation, in a manner that is dependent on the protein's prion-like domain. Temperature controls plant growth and development, and climate change has already altered the phenology of wild plants and crops(1). However, the mechanisms by which plants sense temperature are not well understood. The evening complex is a major signalling hub and a core component of the plant circadian clock(2,3). The evening complex acts as a temperature-responsive transcriptional repressor, providing rhythmicity and temperature responsiveness to growth through unknown mechanisms(2,4-6). The evening complex consists of EARLY FLOWERING 3 (ELF3)(4,7), a large scaffold protein and key component of temperature sensing; ELF4, a small alpha-helical protein; and LUX ARRYTHMO (LUX), a DNA-binding protein required to recruit the evening complex to transcriptional targets. ELF3 contains a polyglutamine (polyQ) repeat(8-10), embedded within a predicted prion domain (PrD). Here we find that the length of the polyQ repeat correlates with thermal responsiveness. We show that ELF3 proteins in plants from hotter climates, with no detectable PrD, are active at high temperatures, and lack thermal responsiveness. The temperature sensitivity of ELF3 is also modulated by the levels of ELF4, indicating that ELF4 can stabilize the function of ELF3. In bothArabidopsisand a heterologous system, ELF3 fused with green fluorescent protein forms speckles within minutes in response to higher temperatures, in a PrD-dependent manner. A purified fragment encompassing the ELF3 PrD reversibly forms liquid droplets in response to increasing temperatures in vitro, indicating that these properties reflect a direct biophysical response conferred by the PrD. The ability of temperature to rapidly shift ELF3 between active and inactive states via phase transition represents a previously unknown thermosensory mechanism.