Abstract

Organisms living in polar waters must cope with an extremely stressful environment dominated by freezing temperatures, high oxygen concentrations and UV radiation. To shed light on the genetic mechanisms on which the polar marine ciliate, Euplotes nobilii, relies to effectively cope with the oxidative stress, attention was focused on methionine sulfoxide reductases which repair proteins with oxidized methionines. A family of four structurally distinct MsrB genes, encoding enzymes specific for the reduction of the methionine-sulfoxide R-forms, were identified from a draft of the E. nobilii transcriptionally active (macronuclear) genome. The En-MsrB genes are constitutively expressed to synthesize proteins markedly different in amino acid sequence, number of CXXC motifs for zinc-ion binding, and presence/absence of a cysteine residue specific for the mechanism of enzyme regeneration. The En-MsrB proteins take different localizations in the nucleus, mitochondria, cytosol and endoplasmic reticulum, ensuring a pervasive protection of all the major subcellular compartments from the oxidative damage. These observations have suggested to regard the En-MsrB gene activity as playing a central role in the genetic mechanism that enables E. nobilii and ciliates in general to live in the polar environment.

Highlights

  • Among the abiotic stresses that affect the life in polar coastal seawaters, high oxygen concentrations and enhanced UV radiations play a central role

  • The amino acid sequence of the previously identified E. nobilii MsrB protein [11], hereafter designated as En-MsrB1, was used as query to search for new genes encoding Msr of type B on the BLAST database of a draft of the E. nobilii macronuclear genome

  • The four En-MsrB genes have the typical organization of the Euplotes macronuclear genes, containing a single open reading frame (ORF) flanked with 5’ leader and 3’ trailer non-coding regions capped with telomeric C4 A4 and G4 T4 repetitions

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Summary

Introduction

Among the abiotic stresses that affect the life in polar coastal seawaters, high oxygen concentrations and enhanced UV radiations play a central role. They are general causes of increased production of reactive oxygen species (ROS). Methionine is one of the most oxidation-sensitive amino acids [1,2,3], and its oxidation by ROS to Met-sulfoxide results in the formation of two, R and S, stereoisomers at the sulfur atom Organisms repair their methionine-oxidized proteins through the enzymatic activity of two structurally distinct classes of methionine sulfoxide reductases, designated as MsrA (EC 1.8.4.11) and MsrB (EC 1.8.4.12). In a previous comparative study between two phylogenetically closely allied ciliate species, namely Euplotes nobilii which is distributed in both Antarctic and Arctic coastal waters [5,6,7,8] and

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