PT - JOURNAL ARTICLE AU - David G. Welkie AU - Benjamin E. Rubin AU - Yong-Gang Chang AU - Spencer Diamond AU - Scott A. Rifkin AU - Andy LiWang AU - Susan S. Golden TI - Genome-wide fitness assessment during diurnal growth reveals an expanded role of the cyanobacterial circadian clock protein KaiA AID - 10.1101/283812 DP - 2018 Jan 01 TA - bioRxiv PG - 283812 4099 - http://biorxiv.org/content/early/2018/06/07/283812.short 4100 - http://biorxiv.org/content/early/2018/06/07/283812.full AB - The recurrent pattern of light and darkness generated by Earth’s axial rotation has profoundly influenced the evolution of organisms, selecting for both biological mechanisms that respond acutely to environmental changes and circadian clocks that program physiology in anticipation of daily variations. The necessity to integrate environmental responsiveness and circadian programming is exemplified in photosynthetic organisms such as cyanobacteria, which depend on light-driven photochemical processes. The cyanobacterium Synechococcus elongatus PCC 7942 is an excellent model system for dissecting these entwined mechanisms. Its core circadian oscillator, consisting of three proteins KaiA, KaiB, and KaiC, transmits time-of-day signals to clock-output proteins, which reciprocally regulate global transcription. Research performed under constant light facilitates analysis of intrinsic cycles separately from direct environmental responses, but does not provide insight into how these regulatory systems are integrated during light-dark cycles. Thus, we sought to identify genes that are specifically necessary in a day-night environment. We screened a dense bar-coded transposon library in both continuous light and daily cycling conditions and compared the fitness consequences of loss of each nonessential gene in the genome. Although the clock itself is not essential for viability in light-dark cycles, the most detrimental mutations revealed by the screen were those that disrupt KaiA. The screen broadened our understanding of light-dark survival in photosynthetic organisms, identified unforeseen clock-protein interaction dynamics, and reinforced the role of the clock as a negative regulator of a night-time metabolic program that is essential for S. elongatus to survive in the dark.Significance Understanding how photosynthetic bacteria respond to and anticipate natural light–dark cycles is necessary for predictive modeling, bioengineering, and elucidating metabolic strategies for diurnal growth. Here, we identify the genetic components that are important specifically under light-dark cycling conditions and determine how a properly functioning circadian clock prepares metabolism for darkness, a starvation period for photoautotrophs. This study establishes that the core circadian clock protein KaiA is necessary to enable rhythmic de-repression of a nighttime circadian program.Specific Contributions DGW BR SD and SSG conceived and designed the project. DGW BR and YGC performed the experiments and analyzed the data. SAR wrote the R scripts for RB-TnSeq conditional fitness analysis. YGC and AL interpreted fluorescence anisotropy data. DGW BR SSG YGC and AL wrote the manuscript.