title: The redox state of SECIS binding protein 2 controls its localization and selenocysteine incorporation function.
abstract: Selenoproteins are central controllers of cellular redox homeostasis. Incorporation of selenocysteine (Sec) into selenoproteins employs a unique mechanism to decode the UGA stop codon. The process requires the Sec insertion sequence (SECIS) element, tRNASec, and protein factors including the SECIS binding protein 2 (SBP2). Here, we report the characterization of motifs within SBP2 that regulate its subcellular localization and function. We show that SBP2 shuttles between the nucleus and the cytoplasm via intrinsic, functional nuclear localization signal and nuclear export signal motifs and that its nuclear export is dependent on the CRM1 pathway. Oxidative stress induces nuclear accumulation of SBP2 via oxidation of cysteine residues within a redox-sensitive cysteine-rich domain. These modifications are efficiently reversed in vitro by human thioredoxin and glutaredoxin, suggesting that these antioxidant systems might regulate redox status of SBP2 in vivo. Depletion of SBP2 in cell lines using small interfering RNA results in a decrease in Sec incorporation, providing direct evidence for its requirement for selenoprotein synthesis. Furthermore, Sec incorporation is reduced substantially after treatment of cells with agents that cause oxidative stress, suggesting that nuclear sequestration of SBP2 under such conditions may represent a mechanism to regulate the expression of selenoproteins.
authors.affiliation: Queensland Institute of Medical Research, 300 Herston Road, Herston, Queensland 4029, Australia.
authors.forename: Laura V
authors.initials: LV
authors.lastname: Papp
authors.forename: Jun
authors.initials: J
authors.lastname: Lu
authors.forename: Frank
authors.initials: F
authors.lastname: Striebel
authors.forename: Derek
authors.initials: D
authors.lastname: Kennedy
authors.forename: Arne
authors.initials: A
authors.lastname: Holmgren
authors.forename: Kum Kum
authors.initials: KK
authors.lastname: Khanna
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D002467:Cell Nucleus / Q000737:chemistry* / Q000378:metabolism; D002478:Cells, Cultured; D003593:Cytoplasm / Q000737:chemistry / Q000378:metabolism; D054477:Glutaredoxins; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D028884:Karyopherins / Q000378:metabolism; D008969:Molecular Sequence Data; D049790:Nuclear Export Signals / Q000235:genetics; D019913:Nuclear Localization Signals / Q000235:genetics / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000378:metabolism; D014176:Protein Biosynthesis; D021381:Protein Transport; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology; D016601:RNA-Binding Proteins / Q000032:analysis* / Q000235:genetics / Q000378:metabolism*; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism; D017279:Selenocysteine / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism; D000097606:Exportin 1 Protein
chemical_list: D054477:Glutaredoxins; D005978:Glutathione; D028884:Karyopherins; D049790:Nuclear Export Signals; D019913:Nuclear Localization Signals; D010088:Oxidoreductases; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D018160:Receptors, Cytoplasmic and Nuclear; D017279:Selenocysteine; D013879:Thioredoxins; D000097606:Exportin 1 Protein; C121530:SECISBP2 protein, human