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Phase separation of GRP7 facilitated by FERONIAmediated phosphorylation inhibits mRNA translation to modulate plant temperature resilience

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Molecular Plant 2024年第3期17卷 460-477页

作者： Fan Xu Long Wang Yingbin Li Junfeng Shi dorothee staiger Feng Yu state Key Laboratory of Chemo/Biosensing and Chemometrics College of Biologyand Hunan Key Laboratory of Plant Functional Genomics and Developmental RegulationHunanUniversityChangsha 410082P.R.China State Key Laboratory of Hybrid Rice Hunan Agricultural Biotechnology Research InstituteHunan Academy of Agricultural SciencesChangsha 410125P.R.China RNA Biology and Molecular Physiology Faculty of BiologyBielefeld University33615 BielefeldGermany

Changes in ambient temperature profoundly affect plant growth and performance.Therefore,the molecu-larbasis of plant acclimation to temperature fluctuation is of great interest.In this study,we discovered that GLYCINE-RICH RNA-BINDING PROTEIN 7(GRP7)contributes to cold and heat tolerance in Arabidopsis thaliana.We found that exposure to a warm temperature rapidly induces GRP7 condensates in planta,which can be reversed by transfer to a lower temperature.Cell biology and biochemical assays revealed that GRP7 undergoes liquid-liquid phase separation(LLPS)in vivo and in vitro.LLPS of GRP7 in the cyto-plasm contributes to the formation of stress granules that recruit RNA,along with the translation machinery component eukaryotic initiation factor 4E1(elF4E1)and the mRNA chaperones COLD SHOCK PROTEIN 1(CSP1)and CSP3,to inhibit translation.Moreover,natural variations in GRP7 affecting the residue phos-phorylated by the receptorkinase FERONIA alter its capacity to undergo LLPS and correlate with the adap-tation of some Arabidopsis accessions to a widertemperature range.Taken together,ourfindings illustrate the role of translational control mediated by GRP7 LLPS to confer plants with temperature resilience.

关键词： fluctuating temperature glycine-rich RNA-binding protein phase separation cold shock proteins translation

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Marking RNA:m^6A writers,readers,and functions in Arabidopsis

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Journal of Molecular Cell Biology 2019年第10期11卷 899-910页

作者： Marlene Reichel Tino Koster dorothee staiger RNA Biology and Molecular Physiology Faculty of BiologyBielefeld University33615 BielefeldGermany

N^6-methyladenosine(m^6A)emerges as an important modification in eukaryotic mRNAs.m^6A has first been reported in 1974,and its functional significance in mammalian gene regulation and importance for proper development have been well established.An arsenal of writer,eraser,and reader proteins accomplish deposition,removal,and interpretation of the m^6A mark,resulting in dynamic function.This led to the concept of an epitranscriptome,the compendium of RNA species with chemical modification ofthe nucleobases in the cell,in analogy to the epigenome.While m^6A has long been known to also exist in plant mRNAs,proteins involved in m^6A metabolism have only recently been detected by mutant analysis,homology search,and mRNA interactome capture in the reference plant Arabidopsis thaliana.Dysregulation ofthe m^6A modification causes severe developmental abnormalities of leaves and roots and altered timing of reproductive development.Furthermore,m^6A modification affects viral infection.Here,we discuss recent progress in identifying m^6A sites transcriptome-wide,in identifying the molecular players involved in writing,removing,and reading the mark,and in assigning functions to this RNA modification in 4.thaliana.We highlight similarities and differences to m^6A modification in mammals and provide an outlook on important questions that remain to be addressed.

关键词： Arabidopsis m^6A mRNA interactome posttranscriptional RNA-binding protein

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A New Set of Reversibly Photoswitchable Fluorescent Proteins for Use in Transgenic Plants

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Molecular Plant 2013年第5期6卷 1518-1530页

作者： Martina Lummer Fabian Humpert Matthias Wiedenubbert Markus Sauer Mark Schuttpelz dorothee staiger Molecular Cell Physiology Bielefeld University Universitatsstr. 25 33615 Bielefeld Germany Institute for Genome Research and Systems Biology CeBiTec 33615 Bielefeld Germany Biomolecular Photonics Bielefeld University Universitatsstr. 25 33615 Bielefeld Germany Biotechnology and Biophysics Julius-Maximilians-University Wurzburg Am Hubland 97074 Wurzburg Germany

Fluorescent reporter proteins that allow repeated switching between a fluorescent and a non-fluorescent state in response to specific wavelengths of light are novel tools for monitoring of protein trafficking and super-resolu- tion fluorescence microscopy in living organisms. Here, we describe variants of the reversibly photoswitchable fluores- cent proteins rsFastLime, bsDronpa, and Padron that have been codon-optimized for the use in transgenic Arabidopsis plants. The synthetic proteins, designated rsFastLIME-s, bsDRONPA-s, and PADRON C-s, showed photophysical properties and switching behavior comparable to those reported for the original proteins. By combining the ＇positively switchable＇ PADRON C-s with the ＇negatively switchable＇ rsFastLIME-s or bsDRONPA-s, two different fluorescent reporter proteins could be imaged at the same wavelength upon transient expression in Nicotiana benthamiana cells. Thus, co-localiza- tion analysis can be performed using only a single detection channel. Furthermore, the proteins were used to tag the RNA-binding protein AtGRP7 （Arabidopsis thaliana glycine-rich RNA-binding protein 7） in transgenic Arabidopsis plants. Because the new reversibly photoswitchable fluorescent proteins show an increase in signal strength during each pho- toactivation cycle, we were able to generate a large number of scans of the same region and reconstruct 3-D images of AtGRP7 expression in the root tip. Upon photoactivation of the AtGRP7：rsFastLIME-s fusion protein in a defined region of a transgenic Arabidopsis root, spreading of the fluorescence signal into adjacent regions was observed, indicating that movement from cell to cell can be monitored. Our results demonstrate that rsFastLIME-s, bsDRONPA-s, and PADRON C-s are versatile fluorescent markers in plants, Furthermore, the proteins also show strong fluorescence in mammalian cells including COS-7 and HeLa cells.

关键词： reversibly photoswitchable fluorescent protein codon usage Arabidopsis rsFastLIME-s bsDRONPA-s PADRON-s.

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