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bioRxiv · 10.1101/2021.03.30.437739

Acclimation of Chlamydomonas reinhardtii to Nitric Oxide Stress Related to Respiratory Burst Oxidase-Like 2

Lee, T.-M.·Kuo, E. Y.
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Abstract

The acclimation mechanism of Chlamydomonas reinhardtii to nitric oxide (NO) was studied by exposure to S-nitroso-N-acetylpenicillamine (SNAP), a NO donor. Treatment with 0.1 or 0.3 mM SNAP transiently inhibited photosynthesis within 1 h, followed by a recovery without growth impairment, while 1.0 mM SNAP treatment caused irreversible photosynthesis inhibition and mortality. The SNAP effects are avoided in the presence of the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide (cPTIO). RNA-seq, qPCR, and biochemical analyses were conducted to decode the metabolic shifts under sub-lethal NO stress by exposure to 0.3 mM SNAP in the presence or absence of 0.4 mM cPTIO. These findings revealed that the acclimation to NO stress comprises a temporally orchestrated implementation of metabolic processes: 1. trigger of NO scavenging elements to reduce NO level; 2. prevention of photo-oxidative risk through photosynthesis inhibition and antioxidant defense system induction; 3. acclimation to nitrogen and sulfur shortage; 4. degradation of damaged proteins through protein trafficking machinery (ubiquitin, SNARE, and autophagy) and molecular chaperone system for dynamic regulation of protein homeostasis. NO increased NADPH oxidase activity and respiratory burst oxidase-like 2 (RBOL2) transcript abundance, which were not observed in the rbol2 insertion mutant. Changes in gene expression in the rbol2 mutant and increased mortality under NO stress demonstrate that NADPH oxidase (RBOL2) is involved in the modulation of some acclimation processes (NO scavenging, antioxidant defense system, autophagy, and heat shock proteins) for Chlamydomonas to cope with NO stress. Our findings provide insight into the molecular events underlying acclimation mechanisms in Chlamydomonas to sub-lethal NO stress. One-sentence SummaryAcclimation machinery is triggered in Chlamydomonas reinhardtii cells against sub-lethal nitric oxide stress.

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Lee, T.-M., Kuo, E. Y.. 2021-03-31. Acclimation of Chlamydomonas reinhardtii to Nitric Oxide Stress Related to Respiratory Burst Oxidase-Like 2. https://doi.org/10.1101/2021.03.30.437739

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