Two Antagonistic Clock-Regulated Histidine Kinases Time the Activation of Circadian Gene Expression

SummaryThe cyanobacterial circadian pacemaker consists of a three-protein clock—KaiA, KaiB, and KaiC—that generates oscillations in the phosphorylation state of KaiC. Here we investigate how temporal information encoded in KaiC phosphorylation is transduced to RpaA, a transcription factor required for circadian gene expression. We show that phosphorylation of RpaA is regulated by two antagonistic histidine kinases, SasA and CikA, which are sequentially activated at distinct times by the Kai clock complex. SasA acts as a kinase toward RpaA, whereas CikA, previously implicated in clock input, acts as a phosphatase that dephosphorylates RpaA. CikA and SasA cooperate to generate an oscillation of RpaA activity that is distinct from that generated by either enzyme alone and offset from the rhythm of KaiC phosphorylation. Our observations reveal how circadian clocks can precisely control the timing of output pathways via the concerted action of two oppositely acting enzymes.

► RpaA is the cognate response regulator of both SasA and CikA
► SasA is the primary kinase for RpaA and is regulated by phosphorylated KaiC
► CikA acts as phosphatase toward RpaA and is regulated by the KaiC/KaiB complex
► RpaA phosphorylation oscillates in vivo, phase advanced to KaiC phosphorylation