In vivo probing of SECIS-dependent selenocysteine translation in Archaea

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

Abstract

Cotranslational insertion of selenocysteine (Sec) proceeds by recoding UGA to a sense codon. This recoding is governed by the Sec insertion sequence (SECIS) element, an RNA structure on the mRNA, but size, location, structure determinants, and mechanism differ for Bacteria, Eukarya, and Archaea. For Archaea, the structure-function relation of the SECIS is poorly understood, as only rather laborious experimental approaches are established. Furthermore, these methods do not allow for quantitative probing of Sec insertion. In order to overcome these limitations, we engineered bacterial β-lactamase into an archaeal selenoprotein, thereby establishing a reporter system, which correlates enzyme activity to Sec insertion. Using this system, in vivo Sec insertion depending on the availability of selenium and the presence of a SECIS element was assessed in Methanococcus maripaludis Furthermore, a minimal SECIS element required for Sec insertion in M. maripaludis was defined and a conserved structural motif shown to be essential for function. Besides developing a convenient tool for selenium research, converting a bacterial enzyme into an archaeal selenoprotein provides proof of concept that novel selenoproteins can be engineered in Archaea.

Details

Original languageEnglish
Article numbere202201676
JournalLife Science Alliance
Volume6
Issue number1
Publication statusPublished - 31 Oct 2022
Peer-reviewedYes

External IDs

Scopus 85140937760
PubMed 36316034
Mendeley d8d7f7fc-6d95-3c52-88b0-b9d7dbe539bc

Keywords

Research priority areas of TU Dresden

Keywords

  • Selenocysteine/genetics, Archaea/genetics, Selenium, 3' Untranslated Regions, Base Sequence, Selenoproteins/genetics

Library keywords