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. 2025 Dec;22(1):1-13.
doi: 10.1080/15476286.2025.2472448. Epub 2025 Mar 2.

Defining the methanogenic SECIS element in vivo by targeted mutagenesis

Nils Peiter  1 , Anna Einert  1 , Pauline Just  1 , Frida Jannasch  1 , Marija Najdovska  1 , Michael Rother  1
Affiliations

Defining the methanogenic SECIS element in vivo by targeted mutagenesis

Nils Peiter et al. RNA Biol. 2025 Dec.

Abstract

In all domains of life, Archaea, Eukarya and Bacteria, the unusual amino acid selenocysteine (Sec) is co-translationally incorporated into proteins by recoding a UGA stop codon to a sense codon. A secondary structure on the mRNA, the selenocysteine insertion sequence (SECIS), is required, but its position, secondary structure and binding partner(s) are not conserved across the tree of life. Thus far, the nature of archaeal SECIS elements has been derived mainly from sequence analyses. A recently developed in vivo reporter system was used to study the structure-function relationships of SECIS elements in Methanococcus maripaludis. Through targeted mutagenesis, we defined the minimal functional SECIS element, the parts of the SECIS where structure and not the identity of the bases are relevant for function, and identified two conserved -and invariant- adenines that are most likely to interact with the other factor(s) of the Sec recoding machinery. Finally, we demonstrated the functionality of SECIS elements in the 5`-untranslated region of the mRNA and identified a potential mechanism of SECIS repositioning in the vicinity of the UGA for efficient selenocysteine insertion.

Keywords: Archaea; Methanococcus; SECIS element; reporter; selenocysteine.

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Conflict of interest statement

No potential conflict of interest was reported by the author(s).

Figures

Figure 1.
Figure 1.
General scheme of the beta-lactamase (Bla) reporter construct.
Figure 2.
Figure 2.
Impact of base exchanges in the basal stem of the methanogenic SECIS element.
Figure 3.
Figure 3.
Impact of base exchanges in the apical stem and terminal loop of the methanogenic SECIS element.
Figure 4.
Figure 4.
Impact of base exchanges in the KTL motif of the methanogenic SECIS element.
Figure 5.
Figure 5.
Abundance of archaeal SECIS elements in the 5-UTR of fdh flanked by coding sequence.
Figure 6.
Figure 6.
SECIS elements in the 5'-UTR.
Figure 7.
Figure 7.
Structure model of the methanogenic SECIS element.
See this image and copyright information in PMC

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