title: The differential expression of glutathione peroxidase 1 and 4 depends on the nature of the SECIS element.
abstract: Selenocysteine insertion into selenoproteins involves the translational recoding of UGA stop codons. In mammals, selenoprotein expression further depends on selenium availability, which has been particularly described for glutathione peroxidase 1 and 4 (Gpx1 and Gpx4). The SECIS element located in the 3'UTR of the selenoprotein mRNAs is a modulator of UGA recoding efficiency in adequate selenium conditions. One of the current models for the UGA recoding mechanism proposes that the SECIS binds SECIS-binding protein 2 (SBP2), which then recruits a selenocysteine-specific elongation factor (EFsec) and tRNA (Sec)  to the ribosome, where L30 acts as an anchor. The involvement of the SECIS in modulation of UGA recoding activity was investigated, together with SBP2 and EFsec, in Hek293 cells cultured with various selenium levels. Luciferase reporter constructs, in transiently or stably expressing cell lines, were used to analyze the differential expression of Gpx1 and Gpx4. We showed that, upon selenium fluctuation, the modulation of UGA recoding efficiency depends on the nature of the SECIS, with Gpx1 being more sensitive than Gpx4. Attenuation of SBP2 and EFsec levels by shRNAs confirmed that both factors are essential for efficient selenocysteine insertion. Strikingly, in a context of either EFsec or SBP2 attenuation, the decrease in UGA recoding efficiency is dependent on the nature of the SECIS, GPx1 being more sensitive. Finally, the profusion of selenium of the culture medium exacerbates the lack of factors involved in selenocysteine insertion.
authors.affiliation: Centre de recherche de Gif-sur-Yvette, FRC 3115, France.
authors.forename: Lynda
authors.initials: L
authors.lastname: Latrèche
authors.forename: Stéphane
authors.initials: S
authors.lastname: Duhieu
authors.forename: Zahia
authors.initials: Z
authors.lastname: Touat-Hamici
authors.forename: Olivier
authors.initials: O
authors.lastname: Jean-Jean
authors.forename: Laurent
authors.initials: L
authors.lastname: Chavatte
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D018388:Codon, Terminator / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D005979:Glutathione Peroxidase / Q000235:genetics* / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D055029:Inverted Repeat Sequences; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010445:Peptide Elongation Factors / Q000235:genetics / Q000378:metabolism; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D014176:Protein Biosynthesis; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012643:Selenium / Q000502:physiology; D000094765:Glutathione Peroxidase GPX1
chemical_list: D020413:3' Untranslated Regions; D018388:Codon, Terminator; C535118:EEFSEC protein, human; D010445:Peptide Elongation Factors; D016601:RNA-Binding Proteins; C121530:SECISBP2 protein, human; D000080662:Phospholipid Hydroperoxide Glutathione Peroxidase; D005979:Glutathione Peroxidase; C523860:glutathione peroxidase 4, rat; D012643:Selenium; D000094765:Glutathione Peroxidase GPX1