ArticlePloS one2017
In vivo and in vitro protein imaging in thermophilic archaea by exploiting a novel protein tag.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 24 citations in OpenAlex.
- Microbial single-cell applications under anoxic conditions.Applied and environmental microbiology · 2024Review
- Archaeal DNA alkylation repair conducted by DNA glycosylase and methyltransferase.Applied microbiology and biotechnology · 2023Review
- A fluorescent reporter system for anaerobic thermophiles.Frontiers in bioengineering and biotechnology · 2023Article
- First thermostable CLIP-Computational and structural biotechnology journal · 2022Article
- The SNAP-Journal of enzyme inhibition and medicinal chemistry · 2021Article
- Live Imaging of a Hyperthermophilic Archaeon Reveals Distinct Roles for Two ESCRT-III Homologs in Ensuring a Robust and Symmetric Division.Current biology : CB · 2020Article
- Review
- Unwinding 20 Years of the Archaeal Minichromosome Maintenance Helicase.Journal of bacteriology · 2020Review
- A journey down to hell: new thermostable protein-tags for biotechnology at high temperatures.Extremophiles : life under extreme conditions · 2020Article
- Establishing Live-Cell Single-Molecule Localization Microscopy Imaging and Single-Particle Tracking in the ArchaeonFrontiers in microbiology · 2020Article
- An AGT-based protein-tag system for the labelling and surface immobilization of enzymes on E. coli outer membrane.Journal of enzyme inhibition and medicinal chemistry · 2019Article
- Multidisciplinary involvement and potential of thermophiles.Folia microbiologica · 2019Review
- Optimization of an In Vitro Transcription/Translation System Based onArchaea (Vancouver, B.C.) · 2019Article
- Every OGT Is Illuminated … by Fluorescent and Synchrotron Lights.International journal of molecular sciences · 2017Review
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Authors and funding
8 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Protein imaging, allowing a wide variety of biological studies both in vitro and in vivo, is of great importance in modern biology. Protein and peptide tags fused to proteins of interest provide the opportunity to elucidate protein location and functions, detect protein-protein interactions, and measure protein activity and kinetics in living cells. Whereas several tags are suitable for protein imaging in mesophilic organisms, the application of this approach to microorganisms living at high temperature has lagged behind. Archaea provide an excellent and unique model for understanding basic cell biology mechanisms. Here, we present the development of a toolkit for protein imaging in the hyperthermophilic archaeon Sulfolobus islandicus. The system relies on a thermostable protein tag (H5) constructed by engineering the alkylguanine-DNA-alkyl-transferase protein of Sulfolobus solfataricus, which can be covalently labeled using a wide range of small molecules. As a suitable host, we constructed, by CRISPR-based genome-editing technology, a S. islandicus mutant strain deleted for the alkylguanine-DNA-alkyl-transferase gene (Δogt). Introduction of a plasmid-borne H5 gene in this strain led to production of a functional H5 protein, which was successfully labeled with appropriate fluorescent molecules and visualized in cell extracts as well as in Δogt live cells. H5 was fused to reverse gyrase, a peculiar thermophile-specific DNA topoisomerase endowed with positive supercoiling activity, and allowed visualization of the enzyme in living cells. To the best of our knowledge, this is the first report of in vivo imaging of any protein of a thermophilic archaeon, filling an important gap in available tools for cell biology studies in these organisms.
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