The use of thermostable fluorescent proteins for live imaging in Sulfolobus acidocaldarius
Abstract: Introduction: Among hyperthermophilic organisms, in vivo protein localization is challenging due to the high growth temperatures that can disrupt proper folding and function of mostly mesophilic-derived fluorescent proteins. While protein localization in the thermophilic model archaeon S. acidocaldarius has been achieved using antibodies with fluorescent probes in fixed cells, the use of thermostable fluorescent proteins for live imaging in thermophilic archaea has so far been unsuccessful. Given the significance of live protein localization in the field of archaeal cell biology, we aimed to identify fluorescent proteins for use in S. acidocaldarius.
Methods: We expressed various previously published and optimized thermostable fluorescent proteins along with fusion proteins of interest and analyzed the cells using flow cytometry and (thermo-) fluorescent microscopy.
Results: Of the tested proteins, thermal green protein (TGP) exhibited the brightest fluorescence when expressed in Sulfolobus cells. By optimizing the linker between TGP and a protein of interest, we could additionally successfully fuse proteins with minimal loss of fluorescence. TGP-CdvB and TGP-PCNA1 fusions displayed localization patterns consistent with previous immunolocalization experiments.
Discussion: These initial results in live protein localization in S. acidocaldarius at high temperatures, combined with recent advancements in thermomicroscopy, open new avenues in the field of archaeal cell biology. This progress finally enables localization experiments in thermophilic archaea, which have so far been limited to mesophilic organisms
- Location
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Deutsche Nationalbibliothek Frankfurt am Main
- Extent
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Online-Ressource
- Language
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Englisch
- Notes
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Frontiers in microbiology. - 15 (2024) , 1445186, ISSN: 1664-302X
- Event
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Veröffentlichung
- (where)
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Freiburg
- (who)
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Universität
- (when)
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2024
- Creator
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Recalde, Alejandra
Abdul-Nabi, Jasmin
Junker, Pierre
Does, Christiaan van der
Elsaesser, Jana
Wolferen, Marleen van
Albers, Sonja Verena
- DOI
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10.3389/fmicb.2024.1445186
- URN
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urn:nbn:de:bsz:25-freidok-2569405
- Rights
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Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Last update
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15.08.2025, 7:21 AM CEST
Data provider
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Associated
- Recalde, Alejandra
- Abdul-Nabi, Jasmin
- Junker, Pierre
- Does, Christiaan van der
- Elsaesser, Jana
- Wolferen, Marleen van
- Albers, Sonja Verena
- Universität
Time of origin
- 2024
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Application of the endogenous CRISPR-Cas type I-D system for genetic engineering in the thermoacidophilic archaeon Sulfolobus acidocaldarius
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Early response of Sulfolobus acidocaldarius to nutrient limitation
Identification of XylR, the activator of Arabinose/Xylose inducible regulon in sulfolobus acidocaldarius and its application for homologous protein expression
Two membrane-bound transcription factors regulate expression of various type-IV-pili surface structures in Sulfolobus acidocaldarius
Optimizing the genetic toolbox of the crenarchaeon Sulfolobus acidocaldarius for use in industrial applications
Adhesion pilus retraction powers twitching motility in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius
SaUspA, the universal stress protein of Sulfolobus acidocaldarius stimulates the activity of the PP2A phosphatase and is involved in growth at high salinity
Putative nucleotide-based second messengers in the archaeal model organisms Haloferax volcanii and Sulfolobus acidocaldarius
AglH, a thermophilic UDP-N-acetylglucosamine-1-phosphate:dolichyl phosphate GlcNAc-1-phosphotransferase initiating protein N-glycosylation pathway in Sulfolobus acidocaldarius, is capable of complementing the eukaryal Alg7
A regulatory RNA is involved in RNA duplex formation and biofilm regulation in Sulfolobus acidocaldarius
AglH, a thermophilic UDP-N-acetylglucosamine-1-phosphate:dolichyl phosphate GlcNAc-1-phosphotransferase initiating protein N-glycosylation pathway in Sulfolobus acidocaldarius, is capable of complementing the eukaryal Alg7
Application of the endogenous CRISPR-Cas type I-D system for genetic engineering in the thermoacidophilic archaeon Sulfolobus acidocaldarius
Role of VapBC4 toxin-antitoxin system of Sulfolobus acidocaldarius in heat stress adaptation
Early response of Sulfolobus acidocaldarius to nutrient limitation
Identification of XylR, the activator of Arabinose/Xylose inducible regulon in sulfolobus acidocaldarius and its application for homologous protein expression
Two membrane-bound transcription factors regulate expression of various type-IV-pili surface structures in Sulfolobus acidocaldarius
Optimizing the genetic toolbox of the crenarchaeon Sulfolobus acidocaldarius for use in industrial applications
Adhesion pilus retraction powers twitching motility in the thermoacidophilic crenarchaeon Sulfolobus acidocaldarius
SaUspA, the universal stress protein of Sulfolobus acidocaldarius stimulates the activity of the PP2A phosphatase and is involved in growth at high salinity
Putative nucleotide-based second messengers in the archaeal model organisms Haloferax volcanii and Sulfolobus acidocaldarius
AglH, a thermophilic UDP-N-acetylglucosamine-1-phosphate:dolichyl phosphate GlcNAc-1-phosphotransferase initiating protein N-glycosylation pathway in Sulfolobus acidocaldarius, is capable of complementing the eukaryal Alg7
A regulatory RNA is involved in RNA duplex formation and biofilm regulation in Sulfolobus acidocaldarius
AglH, a thermophilic UDP-N-acetylglucosamine-1-phosphate:dolichyl phosphate GlcNAc-1-phosphotransferase initiating protein N-glycosylation pathway in Sulfolobus acidocaldarius, is capable of complementing the eukaryal Alg7
Application of the endogenous CRISPR-Cas type I-D system for genetic engineering in the thermoacidophilic archaeon Sulfolobus acidocaldarius
Role of VapBC4 toxin-antitoxin system of Sulfolobus acidocaldarius in heat stress adaptation
Early response of Sulfolobus acidocaldarius to nutrient limitation