Proteins Needed to Activate a Transcriptional Response to the Reactive Oxygen Species Singlet Oxygen

Abstract

ABSTRACT Singlet oxygen ( 1 O 2 ) is a reactive oxygen species generated by energy transfer from one or more excited donors to molecular oxygen. Many biomolecules are prone to oxidation by 1 O 2 , and cells have evolved systems to protect themselves from damage caused by this compound. One way that the photosynthetic bacterium Rhodobacter sphaeroides protects itself from 1 O 2 is by inducing a transcriptional response controlled by ChrR, an anti-σ factor which releases an alternative sigma factor, σ E , in the presence of 1 O 2 . Here we report that induction of σ E -dependent gene transcription is decreased in the presence of 1 O 2 when two conserved genes in the σ E regulon are deleted, including one encoding a cyclopropane fatty acid synthase homologue (RSP2144) or one encoding a protein of unknown function (RSP1091). Thus, we conclude that RSP2144 and RSP1091 are each necessary to increase σ E activity in the presence of 1 O 2 . In addition, we found that unlike in wild-type cells, where ChrR is rapidly degraded when 1 O 2 is generated, turnover of this anti-σ factor is slowed when cells lacking RSP2144, RSP1091, or both of these proteins are exposed to 1 O 2 . Further, we demonstrate that the organic hydroperoxide tert -butyl hydroperoxide promotes ChrR turnover in both wild-type cells and mutants lacking RSP2144 or RSP1091, suggesting differences in the ways different types of oxidants increase σ E activity. IMPORTANCE Oxygen serves many crucial functions on Earth; it is produced during photosynthesis and needed for other pathways. While oxygen is relatively inert, it can be converted to reactive oxygen species (ROS) that destroy biomolecules, cause disease, or kill cells. When energy is transferred to oxygen, the ROS singlet oxygen is generated. To understand how singlet oxygen impacts cells, we study the stress response to this ROS in Rhodobacter sphaeroides , a bacterium that, like plants, generates this compound as a consequence of photosynthesis. This paper identifies proteins that activate a stress response to singlet oxygen and shows that they act in a specific response to this ROS. The identified proteins are found in many free-living, symbiotic, or pathogenic bacteria that can encounter singlet oxygen in nature. Thus, our findings provide new information about a stress response to a ROS of broad biological, agricultural, and biomedical importance.