Selenocysteine tRNA identification in the model organisms Dictyostelium discoideum and Tetrahymena thermophila

Characterizing Sec tRNAs that decode UGA provides one of the most direct and easiest means of determining whether an organism possesses the ability to insert selenocysteine (Sec) into protein. Herein, we used a combination of two techniques, computational to identify Sec tRNA genes and RT-PCR to seq...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2005-04, Vol.329 (1), p.147-151
Main Authors: Shrimali, Rajeev K., Lobanov, Alexey V., Xu, Xue-Ming, Rao, Mahadev, Carlson, Bradley A., Mahadeo, Dana C., Parent, Carole A., Gladyshev, Vadim N., Hatfield, Dolph L.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Computational analysis
Computational Biology
Databases as Topic
Dictyostelium
Dictyostelium - metabolism
Dictyostelium discoideum
Genome
Molecular Sequence Data
Nucleic Acid Conformation
Phylogeny
Protein Structure, Tertiary
Protozoans
Reverse Transcriptase Polymerase Chain Reaction
RNA, Transfer - chemistry
RNA, Transfer, Amino Acid-Specific - chemistry
Selenium
Selenocysteine
Selenocysteine tRNA
Software
Species Specificity
Tetrahymena
Tetrahymena thermophila
Tetrahymena thermophila - metabolism
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Summary:Characterizing Sec tRNAs that decode UGA provides one of the most direct and easiest means of determining whether an organism possesses the ability to insert selenocysteine (Sec) into protein. Herein, we used a combination of two techniques, computational to identify Sec tRNA genes and RT-PCR to sequence the gene products, to unequivocally demonstrate that two widely studied, model protozoans, Dictyostelium discoideum and Tetrahymena thermophila, encode Sec tRNA in their genomes. The advantage of using both procedures is that computationally we could easily detect potential Sec tRNA genes and then confirm by sequencing that the Sec tRNA was present in the tRNA population, and thus the identified gene was not a pseudogene. Sec tRNAs from both organisms decode UGA. T. thermophila Sec tRNA, like all other sequenced Sec tRNAs, is 90 nucleotides in length, while that from D. discoideum is 91 nucleotides long making it the longest eukaryotic sequenced to date. Evolutionary analyses of known Sec tRNAs reveal the two forms identified herein are the most divergent eukaryotic Sec tRNAs thus far sequenced.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2005.01.120