ArticleNucleic acids research2003
Revised Escherichia coli selenocysteine insertion requirements determined by in vivo screening of combinatorial libraries of SECIS variants.
Article in Nucleic acids research, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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10 citing papers in PubMed, 27 citations in OpenAlex.
- RNAiFold 2.0: a web server and software to design custom and Rfam-based RNA molecules.Nucleic acids research · 2015Article
- Multivalent site-specific phage modification enhances the binding affinity of receptor ligands.Bioconjugate chemistry · 2015Article
- Tandem use of selenocysteine: adaptation of a selenoprotein glutaredoxin for reduction of selenoprotein methionine sulfoxide reductase.Molecular microbiology · 2011Article
- In-depth genetic analysis of Clostridium difficile PCR-ribotype 027 strains reveals high genome fluidity including point mutations and inversions.Gut microbes · 2010Article
- Proteomic profiling of L-cysteine induced selenite resistance in Enterobacter sp. YSU.Proteome science · 2009Article
- Probing the role of the proximal heme ligand in cytochrome P450cam by recombinant incorporation of selenocysteine.Proceedings of the National Academy of Sciences of the United States of America · 2009Article
- The selenoproteome of Clostridium sp. OhILAs: characterization of anaerobic bacterial selenoprotein methionine sulfoxide reductase A.Proteins · 2009Article
- Factors and selenocysteine insertion sequence requirements for the synthesis of selenoproteins from a gram-positive anaerobe in Escherichia coli.Applied and environmental microbiology · 2008Article
- Comparative context analysis of codon pairs on an ORFeome scale.Genome biology · 2005Article
- Selenocysteine insertion directed by the 3'-UTR SECIS element in Escherichia coli.Nucleic acids research · 2005Article
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4 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
To investigate the stringency of the Escherichia coli selenocysteine insertion sequence (SECIS) requirements, libraries of SECIS variants were screened via a novel method in which suppression of the selenocysteine (Sec) opal codon was coupled to bacteriophage plaque formation. The SECIS variant libraries were designed with a mostly paired lower stem, so that randomization could be focused on the upper stem and loop regions. We identified 19 functional non-native SECIS sequences that violated the expected pairing requirements for the SECIS upper stem. All of the SECIS variants were shown to permit Sec insertion in phage (by chemical modification of the Sec residue) and fused to lacZalpha (by beta-galactosidase assay). The diminished pairing of the upper stem appears to be mitigated by the overall stem stability; a given upper stem variant has significantly higher readthrough in the context of a paired, rather than unpaired, lower stem. These results suggest an unexpected downstream sequence flexibility in prokaryotic selenoprotein expression.
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