Time-of-day-dependent responses of cyanobacterial cellular viability against oxidative stress

Kenya Tanaka,Shuji Nakanishi,Ginga Shimakawa

doi:10.1038/s41598-020-77141-8

Abstract

As an adaptation to periodic fluctuations of environmental light, photosynthetic organisms have evolved a circadian clock. Control by the circadian clock of many cellular physiological functions, including antioxidant enzymes, metabolism and the cell cycle, has attracted attention in the context of oxidative stress tolerance. However, since each physiological function works in an integrated manner to deal with oxidative stress, whether or not cell responses to oxidative stress are under circadian control remains an open question. In fact, circadian rhythms of oxidative stress tolerance have not yet been experimentally demonstrated. In the present work, we applied an assay using methyl viologen (MV), which generates reactive oxygen species (ROS) under light irradiation, and experimentally verified the circadian rhythms of oxidative stress tolerance in photosynthetic cells of the cyanobacterium Synechococcus elongatus PCC 7942, a standard model species for investigation of the circadian clock. Here, we report that ROS generated by MV treatment causes damage to stroma components and not to the photosynthetic electron transportation chain, leading to reduced cell viability. The degree of decrease in cell viability was dependent on the subjective time at which oxidative stress was applied. Thus, oxidative stress tolerance was shown to exhibit circadian rhythms. In addition, the rhythmic pattern of oxidative stress tolerance disappeared in mutant cells lacking the essential clock genes. Notably, ROS levels changed periodically, independent of the MV treatment. Thus, we demonstrate for the first time that in cyanobacterial cells, oxidative stress tolerance shows circadian oscillation.

Highlights

As an adaptation to periodic fluctuations of environmental light, photosynthetic organisms have evolved a circadian clock
S. elongatus cells grown in a turbidostat were assayed using a protocol in which 50 μM methyl viologen (MV) was added to cell culture aliquots, followed by 30 min light irradiation
After MV/light-treatment, cells were washed with BG-11 medium and oxidative damage was evaluated by measuring oxygen evolution activity, P700 absorbance change or colony forming units (CFU) (Fig. 1)

Summary

Introduction

As an adaptation to periodic fluctuations of environmental light, photosynthetic organisms have evolved a circadian clock. Since daily fluctuations in environmental light levels lead to periodic variations in reactive oxygen species (ROS) levels in photosynthetic organisms, the physiological function of the circadian clock has been thought to be relevant to oxidative stress tolerance. The cyanobacterium Synechococcus elongatus PCC 7942 (hereafter, S. elongatus), a photosynthetic autotrophic bacterium, is an appropriate model for study of circadian control of cellular physiology because the molecular mechanisms underlying its circadian clock is well understood, as summarized in recent reviews[4,5] In this organisms, attention has been paid to relationship between circadian clock and oxidative stress. In photosynthetic living cells, the generation of large amounts of ROS is triggered by unfavorable conditions such as strong light irradiation or exposure to high (or low) temperature These environmental stimuli may mask the circadian control of oxidative stress tolerance because they directly affect a variety of metabolic pathways and induce adaptation mechanisms against oxidative stress[10]. Circadian rhythms of cell viability were observed when MV treatment with relatively low light intensity was used

Methods

Results

Conclusion