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Circadian clocks are biochemical oscillatory mechanisms that co-ordinate with a 24h period predicting the daily diurnal changes in the environment to regulate various metabolic, developmental and physiological processes in the organisms. The plant circadian clock represents a network of interconnected multiple feedback loops at the core of which lie two Myb-like transcription factors, Circadian Clock Associated 1 (CCA1) and Late Elongated Hypocotyl (LHY), that express in the morning, and an evening expressing Timing of CAB Expression 1 (TOC1). Other components important for the clock are Early Flowering 3 and 4 (ELF3 and ELF4) and Lux Arrhythmo (LUX) that accumulate in the evening (Harmer, 2009). Previous studies have shown these to be part of an evening complex that is important for the regulation of the core oscillator (Nusinow et al., 2011; Herrero et al., 2012). Loss-of-function mutation studies of these components demonstrate arrhythmia under constant conditions of light (LL) and in darkness (DD) (Kolmos et al., 2009; Thines and Harmon, 2010). Recently, it has been shown (both in vivo and in vitro) that ELF3 and ELF4 interact physically with each other to perform their regulatory function (Herrero et al., 2012). Here, using various in silico and experimental approaches, we have tried to understand the structural aspects of these two evening expressing plant clock proteins (ELF3 and 4). Due to the instability of the ELF4 and ELF3 proteins, crystallization and Small Angle X-ray Scattering (SAXS) measurements were carried out for recombinant variants tagged with fusion (MBP or NusA) proteins.

Biochemical and structural studies of plant circadian clock proteins