Chapter title |
Selenocysteine tRNA[Ser]Sec, the Central Component of Selenoprotein Biosynthesis: Isolation, Identification, Modification, and Sequencing
|
---|---|
Chapter number | 4 |
Book title |
Selenoproteins
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7258-6_4 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7257-9, 978-1-4939-7258-6
|
Authors |
Bradley A. Carlson, Byeong Jae Lee, Petra A. Tsuji, Paul R. Copeland, Ulrich Schweizer, Vadim N. Gladyshev, Dolph L. Hatfield |
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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