Abstract
The selenocysteine (Sec) tRNA[Ser]Sec population consists of two isoforms that differ from each other by a single 2′-O-methylribosyl moiety at position 34 (Um34). These two isoforms, which are encoded in a single gene, Trsp, and modified posttranscriptionally, are involved individually in the synthesis of two subclasses of selenoproteins, designated housekeeping and stress-related selenoproteins. Techniques used in obtaining these isoforms for their characterization include extraction of RNA from mammalian cells and tissues, purifying the tRNA[Ser]Sec population by one or more procedures, and finally resolving the two isoforms from each other. Since some of the older techniques for isolating tRNA[Ser]Sec and resolving the isoforms are used in only a few laboratories, these procedures will be discussed briefly and references provided for more detailed information, while the more recently developed procedures are discussed in detail. In addition, a novel technique that was developed in sequencing tRNA[Ser]Sec for identifying their occurrence in other organisms is also presented.
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Acknowledgments
This work was supported by the Intramural Research Program of the National Institutes of Health, NCI, Center for Cancer Research to DLH, NIH grants CA080946, GM061603 and GM065204 to VNG, DFG Priority Program 1784 to US (Schw914/5-1) to US, Towson University’s Jess and Mildred Fisher College of Science and Mathematics to PAT, who is a Jess and Mildred Fisher Endowed Chair of Biological Sciences, and NIH grants GM077073 and HD083616 to PRC.
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Carlson, B.A. et al. (2018). Selenocysteine tRNA[Ser]Sec, the Central Component of Selenoprotein Biosynthesis: Isolation, Identification, Modification, and Sequencing. In: Chavatte, L. (eds) Selenoproteins. Methods in Molecular Biology, vol 1661. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7258-6_4
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DOI: https://doi.org/10.1007/978-1-4939-7258-6_4
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