Novel selenoproteins identified in silico and in vivo by using a conserved RNA structural motif.
Selenocysteine is incorporated into selenoproteins by an in-frame UGA codon whose readthrough requires the selenocysteine insertion sequence (SECIS), a conserved hairpin in the 3'-untranslated region of eukaryotic selenoprotein mRNAs. To identify new selenoproteins, we developed a strategy that obviates the need for prior amino acid sequence information. A computational screen was used to scan nucleotide sequence data bases for sequences presenting a potential SECIS secondary structure. The computer-selected hairpins were then assayed in vivo for their functional capacities, and the cDNAs corresponding to the SECIS winners were identified. Four of them encoded novel selenoproteins as confirmed by in vivo experiments. Among these, SelZf1 and SelZf2 share a common domain with mitochondrial thioredoxin reductase-2. The three proteins, however, possess distinct N-terminal domains. We found that another protein, SelX, displays sequence similarity to a protein involved in bacterial pilus formation. For the first time, four novel selenoproteins were discovered based on a computational screen for the RNA hairpin directing selenocysteine incorporation.
Lescure
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Unité Propre de Recherche CNRS 9002, Structure des Macromolécules Biologiques et Mécanismes de Reconnaissance, Institut de Biologie Moleculaire et Cellulaire, 15, Rue René Descartes, 67084 Strasbourg Cedex, France.
Gautheret
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Carbon
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Krol
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D019295:Computational Biology; D017124:Conserved Sequence; D018076:DNA, Complementary; D006801:Humans; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011506:Proteins / Q000737:chemistry*; D012313:RNA / Q000737:chemistry*; D017279:Selenocysteine / Q000737:chemistry; D051140:Selenoproteins; D017386:Sequence Homology, Amino Acid
D003062:Codon; D018076:DNA, Complementary; D011506:Proteins; D051140:Selenoproteins; D017279:Selenocysteine; D012313:RNA