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
 
Download
Es wurden keine Dateien hochgeladen. Nur Publikationsnachweis!
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. https://doi.org/10.1111/j.1600-0854.2011.01180.x
Lummer, Martina, Humpert, Fabian, Steuwe, Christian, Caesar, Katharina, Schüttpelz, Mark, Sauer, Markus, and Staiger, Dorothee. 2011. “Reversible Photoswitchable DRONPA-s Monitors Nucleocytoplasmic Transport of an RNA-Binding Protein in Transgenic Plants”. Traffic 12 (6): 693-702.
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.

20 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Advances in Imaging Plant Cell Dynamics.
Komis G, Novák D, Ovečka M, Šamajová O, Šamaj J., Plant Physiol 176(1), 2018
PMID: 29167354
Photoinducible DRONPA-s: a new tool for investigating cell-cell connectivity.
Gerlitz N, Gerum R, Sauer N, Stadler R., Plant J 94(5), 2018
PMID: 29654648
Beyond Transcription: Fine-Tuning of Circadian Timekeeping by Post-Transcriptional Regulation.
Mateos JL, de Leone MJ, Torchio J, Reichel M, Staiger D., Genes (Basel) 9(12), 2018
PMID: 30544736
Transient plant transformation mediated by Agrobacterium tumefaciens: Principles, methods and applications.
Krenek P, Samajova O, Luptovciak I, Doskocilova A, Komis G, Samaj J., Biotechnol Adv 33(6 pt 2), 2015
PMID: 25819757
Salicylic acid-dependent and -independent impact of an RNA-binding protein on plant immunity.
Hackmann C, Korneli C, Kutyniok M, Köster T, Wiedenlübbert M, Müller C, Staiger D., Plant Cell Environ 37(3), 2014
PMID: 23961939
A glycine-rich RNA-binding protein affects gibberellin biosynthesis in Arabidopsis.
Löhr B, Streitner C, Steffen A, Lange T, Staiger D., Mol Biol Rep 41(1), 2014
PMID: 24281950
HnRNP-like proteins as post-transcriptional regulators.
Yeap WC, Namasivayam P, Ho CL., Plant Sci 227(), 2014
PMID: 25219311
Regulation of pri-miRNA processing by the hnRNP-like protein AtGRP7 in Arabidopsis.
Köster T, Meyer K, Weinholdt C, Smith LM, Lummer M, Speth C, Grosse I, Weigel D, Staiger D., Nucleic Acids Res 42(15), 2014
PMID: 25104024
Mutational definition of binding requirements of an hnRNP-like protein in Arabidopsis using fluorescence correlation spectroscopy.
Leder V, Lummer M, Tegeler K, Humpert F, Lewinski M, Schüttpelz M, Staiger D., Biochem Biophys Res Commun 453(1), 2014
PMID: 25251471
Emerging role for RNA-based regulation in plant immunity.
Staiger D, Korneli C, Lummer M, Navarro L., New Phytol 197(2), 2013
PMID: 23163405
Elements of transcriptional machinery are compatible among plants and mammals.
Wolf A, Akrap N, Marg B, Galliardt H, Heiligentag M, Humpert F, Sauer M, Kaltschmidt B, Kaltschmidt C, Seidel T., PLoS One 8(1), 2013
PMID: 23326494
Pseudomonas HopU1 modulates plant immune receptor levels by blocking the interaction of their mRNAs with GRP7.
Nicaise V, Joe A, Jeong BR, Korneli C, Boutrot F, Westedt I, Staiger D, Alfano JR, Zipfel C., EMBO J 32(5), 2013
PMID: 23395902
A new set of reversibly photoswitchable fluorescent proteins for use in transgenic plants.
Lummer M, Humpert F, Wiedenlübbert M, Sauer M, Schüttpelz M, Staiger D., Mol Plant 6(5), 2013
PMID: 23434876
Localization and dynamics of nuclear speckles in plants.
Reddy AS, Day IS, Göhring J, Barta A., Plant Physiol 158(1), 2012
PMID: 22045923
An hnRNP-like RNA-binding protein affects alternative splicing by in vivo interaction with transcripts in Arabidopsis thaliana.
Streitner C, Köster T, Simpson CG, Shaw P, Danisman S, Brown JW, Staiger D., Nucleic Acids Res 40(22), 2012
PMID: 23042250

42 References

Daten bereitgestellt von Europe PubMed Central.

RNA-binding proteins and post-transcriptional gene regulation.
Glisovic T, Bachorik JL, Yong J, Dreyfuss G., FEBS Lett. 582(14), 2008
PMID: 18342629
AtGRP7, a nuclear RNA-binding protein as a component of a circadian-regulated negative feedback loop in Arabidopsis thaliana.
Heintzen C, Nater M, Apel K, Staiger D., Proc. Natl. Acad. Sci. U.S.A. 94(16), 1997
PMID: 9238008
The small glycine-rich RNA binding protein AtGRP7 promotes floral transition in Arabidopsis thaliana.
Streitner C, Danisman S, Wehrle F, Schoning JC, Alfano JR, Staiger D., Plant J. 56(2), 2008
PMID: 18573194
Global transcript profiling of transgenic plants constitutively overexpressing the RNA-binding protein AtGRP7.
Streitner, BMC Plant Biol 10(), 2010
A type III effector ADP-ribosylates RNA-binding proteins and quells plant immunity.
Fu ZQ, Guo M, Jeong BR, Tian F, Elthon TE, Cerny RL, Staiger D, Alfano JR., Nature 447(7142), 2007
PMID: 17450127
Auto-regulation of the circadian slave oscillator component AtGRP7 and regulation of its targets is impaired by a single RNA recognition motif point mutation.
Schoning JC, Streitner C, Page DR, Hennig S, Uchida K, Wolf E, Furuya M, Staiger D., Plant J. 52(6), 2007
PMID: 17924945
Changes in conformational dynamics of mRNA upon AtGRP7 binding studied by fluorescence correlation spectroscopy.
Schuttpelz M, Schoning JC, Doose S, Neuweiler H, Peters E, Staiger D, Sauer M., J. Am. Chem. Soc. 130(29), 2008
PMID: 18576621
Transport between the cell nucleus and the cytoplasm.
Gorlich D, Kutay U., Annu. Rev. Cell Dev. Biol. 15(), 1999
PMID: 10611974
Characterisation of the passive permeability barrier of nuclear pore complexes.
Mohr D, Frey S, Fischer T, Guttler T, Gorlich D., EMBO J. 28(17), 2009
PMID: 19680228
Mechanisms and signals for the nuclear import of proteins.
Freitas N, Cunha C., Curr. Genomics 10(8), 2009
PMID: 20514217
A nuclear localization domain in the hnRNP A1 protein.
Siomi H, Dreyfuss G., J. Cell Biol. 129(3), 1995
PMID: 7730395
Definition of a consensus transportin-specific nucleocytoplasmic transport signal.
Bogerd HP, Benson RE, Truant R, Herold A, Phingbodhipakkiya M, Cullen BR., J. Biol. Chem. 274(14), 1999
PMID: 10092666
Arabidopsis transportin1 is the nuclear import receptor for the circadian clock-regulated RNA-binding protein AtGRP7.
Ziemienowicz A, Haasen D, Staiger D, Merkle T., Plant Mol. Biol. 53(1-2), 2003
PMID: 14756317
Glycine-rich RNA-binding protein 7 affects abiotic stress responses by regulating stomata opening and closing in Arabidopsis thaliana.
Kim JS, Jung HJ, Lee HJ, Kim KA, Goh CH, Woo Y, Oh SH, Han YS, Kang H., Plant J. 55(3), 2008
PMID: 18410480
Regulated fast nucleocytoplasmic shuttling observed by reversible protein highlighting.
Ando R, Mizuno H, Miyawaki A., Science 306(5700), 2004
PMID: 15550670
The fluorescence dynamics of single molecules of green fluorescent protein.
Peterman, J Phys Chem 103(), 1999
Exo- and endocytotic trafficking of SCAMP2.
Toyooka K, Matsuoka K., Plant Signal Behav 4(12), 2009
PMID: 20514246
Functional distribution and dynamics of Arabidopsis SR splicing factors in living plant cells.
Tillemans V, Dispa L, Remacle C, Collinge M, Motte P., Plant J. 41(4), 2005
PMID: 15686520
Insights into nuclear organization in plants as revealed by the dynamic distribution of Arabidopsis SR splicing factors.
Tillemans V, Leponce I, Rausin G, Dispa L, Motte P., Plant Cell 18(11), 2006
PMID: 17114353
Analyses of in vivo interaction and mobility of two spliceosomal proteins using FRAP and BiFC.
Ali, PLoS One 3(), 2008
Dynamic nucleocytoplasmic shuttling of an Arabidopsis SR splicing factor: role of the RNA-binding domains.
Rausin G, Tillemans V, Stankovic N, Hanikenne M, Motte P., Plant Physiol. 153(1), 2010
PMID: 20237019
Glycine-rich RNA-binding protein 7 affects abiotic stress responses by regulating stomata opening and closing in Arabidopsis thaliana.
Kim JS, Jung HJ, Lee HJ, Kim KA, Goh CH, Woo Y, Oh SH, Han YS, Kang H., Plant J. 55(3), 2008
PMID: 18410480
The role of the plant nucleolus in pre-mRNA processing.
Brown JW, Shaw PJ., Curr. Top. Microbiol. Immunol. 326(), 2008
PMID: 18630759
Mouse period 2 mRNA circadian oscillation is modulated by PTB-mediated rhythmic mRNA degradation.
Woo KC, Kim TD, Lee KH, Kim DY, Kim W, Lee KY, Kim KT., Nucleic Acids Res. 37(1), 2008
PMID: 19010962
Spotlight on post-transcriptional control in the circadian system.
Staiger, Cell Mol Life Sci 68(), 2010
In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants.
Bechtold, Life Science de la vie/Life Sciences 316(), 1993
A revised medium for rapid growth and bio assays with tobacco tissue cultures.
Murashige, Physiol Plantarum 15(), 1962
A set of plant expression vectors for transcriptional and translational fusions.
Töpfer, Nucl Acids Res 15(), 1987
The promotor of TL-DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type of Agrobacterium binary vector.
Koncz, Mol Gen Genet 204(), 1986
Export

Markieren/ Markierung löschen
Markierte Publikationen

Open Data PUB

Web of Science

Dieser Datensatz im Web of Science®
Quellen

PMID: 21453442
PubMed | Europe PMC

Suchen in

Google Scholar