Selenocysteine tRNA[Ser]Sec, the central component of selenoprotein biosynthesis: Isolation, identification, modification, and sequencing

Bradley A. Carlson, Byeong Jae Lee, Petra A. Tsuji, Paul R. Copeland, Ulrich Schweizer, Vadim N. Gladyshev, Dolph L. Hatfield

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages43-60
Number of pages18
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1661
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Chromatography
  • Gel electrophoresis
  • Selenium
  • Selenocysteine tRNA
  • Selenocysteine tRNA detection
  • Selenocysteine tRNA gene modification

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    Carlson, B. A., Lee, B. J., Tsuji, P. A., Copeland, P. R., Schweizer, U., Gladyshev, V. N., & Hatfield, D. L. (2018). Selenocysteine tRNA[Ser]Sec, the central component of selenoprotein biosynthesis: Isolation, identification, modification, and sequencing. In Methods in Molecular Biology (pp. 43-60). (Methods in Molecular Biology; Vol. 1661). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7258-6_4