Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants

Lummer M, Humpert F, Steuwe C, Caesar K, Schüttpelz M, Sauer M, Staiger D (2011)
Traffic 12(6): 693-702.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Abstract / Bemerkung
Fluorescent reporter proteins that allow repeated switching between a fluorescent and a non-fluorescent state are novel tools for monitoring intracellular protein trafficking. A codon-optimized variant of the reversibly photoswitchable fluorescent protein DRONPA was designed for the use in transgenic Arabidopsis plants. Its codon usage is also well adapted to the mammalian codon usage. The synthetic protein, DRONPA-s, shows photochemical properties and switching behavior comparable to that of the original DRONPA from Pectiniidae both in vitro and in vivo. DRONPA-s fused to the RNA-binding protein AtGRP7 (Arabidopsis thaliana glycine-rich RNA-binding protein 7) under control of the endogenous AtGRP7 promoter localizes to cytoplasm, nucleoplasm and nucleolus of transgenic Arabidopsis plants. To monitor the intracellular transport dynamics of AtGRP7-DRONPA-s, we set up a single-molecule sensitive confocal fluorescence microscope. Fluorescence recovery after selective photoswitching experiments revealed that AtGRP7-DRONPA-s reaches the nucleus by carrier-mediated transport. Furthermore, photoactivation experiments showed that AtGRP7-DRONPA-s is exported from the nucleus. Thus, AtGRP7 is a nucleocytoplasmic shuttling protein. Our results show that the fluorescent marker DRONPA-s is a versatile tool to track protein transport dynamics in stably transformed plants.
Stichworte
reversible photoswitchable; Arabidopsis; codon usage; fluorescent protein; DRONPA; export; selective photoswitching; nuclear import; fluorescence recovery after; RNA-binding protein
Erscheinungsjahr
2011
Zeitschriftentitel
Traffic
Band
12
Ausgabe
6
Seite(n)
693-702
ISSN
1398-9219
Page URI
https://pub.uni-bielefeld.de/record/2289841

Zitieren

Lummer M, Humpert F, Steuwe C, et al. Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants. Traffic. 2011;12(6):693-702.
Lummer, M., Humpert, F., Steuwe, C., Caesar, K., Schüttpelz, M., Sauer, M., & Staiger, D. (2011). Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants. Traffic, 12(6), 693-702. doi:10.1111/j.1600-0854.2011.01180.x
Lummer, M., Humpert, F., Steuwe, C., Caesar, K., Schüttpelz, M., Sauer, M., and Staiger, D. (2011). Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants. Traffic 12, 693-702.
Lummer, M., et al., 2011. Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants. Traffic, 12(6), p 693-702.
M. Lummer, et al., “Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants”, Traffic, vol. 12, 2011, pp. 693-702.
Lummer, M., Humpert, F., Steuwe, C., Caesar, K., Schüttpelz, M., Sauer, M., Staiger, D.: Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants. Traffic. 12, 693-702 (2011).
Lummer, Martina, Humpert, Fabian, Steuwe, Christian, Caesar, Katharina, Schüttpelz, Mark, Sauer, Markus, and Staiger, Dorothee. “Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants”. Traffic 12.6 (2011): 693-702.

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