Selenocysteine, a selenium-containing analog of cysteine, is found in the prokaryotic and eukaryotic kingdoms in active sites of enzymes involved in oxidation-reduction reactions. This aminoacid is cotranslationally i...Selenocysteine, a selenium-containing analog of cysteine, is found in the prokaryotic and eukaryotic kingdoms in active sites of enzymes involved in oxidation-reduction reactions. This aminoacid is cotranslationally incorporated at UGA codons which usually act as translation stop codons. In eukaryotes, decoding of selenocysteine necessitates the participation of the selenocysteine insertion sequence (SECIS), an element lying in the 3' -untranslated region of selenoprotein mRNAs. A detailed experimental study of the secondary structures of the SECIS elements of rat and human type 1 iodothyronine deiodinases and rat glutathione peroxidase was performed. Enzymatic and chemical structure probing led us to propose a secondary structure model, supported by sequence comparison of 23 SECIS mRNAs. The secondary structure model revealed the existence of a novel type of RNA motif composed of four consecutive non-Watson-Crick base-pairs. Using gel shift experiments, we identified in several mammalian cell type extracts the protein SBP,for SECIS-binding protein, that specifically recognizes the iodothyronine deiodinases and glutathione peroxidase SECIS elements. The structural model that we derived for the SECIS RNAs discloses RNA features possibly implicated in the binding of SBP and/or SECIS function展开更多
Incorporation of Selenocysteine into protein requires an RNA structural motif, SECIS (Selenocysteine insertion sequence) element that, along with other factors, demarcates UGA-Sec from the UGA termination codon, for e...Incorporation of Selenocysteine into protein requires an RNA structural motif, SECIS (Selenocysteine insertion sequence) element that, along with other factors, demarcates UGA-Sec from the UGA termination codon, for expression of Selenoproteins (in case of eukaryotes). It has been predicted that during HIV infection, several functional viral selenoproteins are expressed and synthesis of these viral selenoproteins deplete the selenium level of the host. It might be that even the viral genome has the SECIS elements in their Selenoprotein mRNA, and during infection, the host cellular machinery is transformed in such a way that the human Sec tRNA binds to the viral Selenoprotein mRNA, instead of binding to its own Selenoprotein mRNA, thus leading to expression of viral selenoproteins. This hypothesis was tested in this study by identifying the SECIS elements in the HIV-1 genome and further predicting their secondary and tertiary structures. We then tried to dock these tertiary structures with human Sec tRNA. Here we report putatively the presence of 3215 SECIS elements in the HIV-1 genome and that the human Sec tRNAsec binds to the viral SECIS elements present in the viral selenoprotein mRNA. Based on an earlier finding, it was observed that atoms of A8 and U9, which present in human Sec tRNA, are the possible key sites for binding.展开更多
The computer program RNA Draw was used to identify the secondary structures in the 3’untranslated regions (3’UTRs) of the mRNAs from 46 eukaryotic seleno-proteins among 7 species. The program found one or two possib...The computer program RNA Draw was used to identify the secondary structures in the 3’untranslated regions (3’UTRs) of the mRNAs from 46 eukaryotic seleno-proteins among 7 species. The program found one or two possible SECIS elements in these selenoproteins. The SECIS element consists of a stem-loop or hairpin structure with three conserved sequences of AUGA-(A)AA-GA. SECIS element was not found by the RNA Draw program in randomly selected non-selenoproteins. The results showed that SECIS element is the unique character of the genes ofeukaryotic selenoproteins. Thus it is possible to use RNA Draw to search the SECIS elements in gene bank for potential new selenoproteins.展开更多
文摘Selenocysteine, a selenium-containing analog of cysteine, is found in the prokaryotic and eukaryotic kingdoms in active sites of enzymes involved in oxidation-reduction reactions. This aminoacid is cotranslationally incorporated at UGA codons which usually act as translation stop codons. In eukaryotes, decoding of selenocysteine necessitates the participation of the selenocysteine insertion sequence (SECIS), an element lying in the 3' -untranslated region of selenoprotein mRNAs. A detailed experimental study of the secondary structures of the SECIS elements of rat and human type 1 iodothyronine deiodinases and rat glutathione peroxidase was performed. Enzymatic and chemical structure probing led us to propose a secondary structure model, supported by sequence comparison of 23 SECIS mRNAs. The secondary structure model revealed the existence of a novel type of RNA motif composed of four consecutive non-Watson-Crick base-pairs. Using gel shift experiments, we identified in several mammalian cell type extracts the protein SBP,for SECIS-binding protein, that specifically recognizes the iodothyronine deiodinases and glutathione peroxidase SECIS elements. The structural model that we derived for the SECIS RNAs discloses RNA features possibly implicated in the binding of SBP and/or SECIS function
文摘Incorporation of Selenocysteine into protein requires an RNA structural motif, SECIS (Selenocysteine insertion sequence) element that, along with other factors, demarcates UGA-Sec from the UGA termination codon, for expression of Selenoproteins (in case of eukaryotes). It has been predicted that during HIV infection, several functional viral selenoproteins are expressed and synthesis of these viral selenoproteins deplete the selenium level of the host. It might be that even the viral genome has the SECIS elements in their Selenoprotein mRNA, and during infection, the host cellular machinery is transformed in such a way that the human Sec tRNA binds to the viral Selenoprotein mRNA, instead of binding to its own Selenoprotein mRNA, thus leading to expression of viral selenoproteins. This hypothesis was tested in this study by identifying the SECIS elements in the HIV-1 genome and further predicting their secondary and tertiary structures. We then tried to dock these tertiary structures with human Sec tRNA. Here we report putatively the presence of 3215 SECIS elements in the HIV-1 genome and that the human Sec tRNAsec binds to the viral SECIS elements present in the viral selenoprotein mRNA. Based on an earlier finding, it was observed that atoms of A8 and U9, which present in human Sec tRNA, are the possible key sites for binding.
文摘The computer program RNA Draw was used to identify the secondary structures in the 3’untranslated regions (3’UTRs) of the mRNAs from 46 eukaryotic seleno-proteins among 7 species. The program found one or two possible SECIS elements in these selenoproteins. The SECIS element consists of a stem-loop or hairpin structure with three conserved sequences of AUGA-(A)AA-GA. SECIS element was not found by the RNA Draw program in randomly selected non-selenoproteins. The results showed that SECIS element is the unique character of the genes ofeukaryotic selenoproteins. Thus it is possible to use RNA Draw to search the SECIS elements in gene bank for potential new selenoproteins.
