TY - JOUR A1 - Ralevski, Alexandra A1 - Apelt, Federico A1 - Olas, Justyna Jadwiga A1 - Müller-Röber, Bernd A1 - Rugarli, Elena I. A1 - Kragler, Friedrich A1 - Horvath, Tamas L. T1 - Plant mitochondrial FMT and its mammalian homolog CLUH controls development and behavior in Arabidopsis and locomotion in mice JF - Cellular and molecular life sciences N2 - Mitochondria in animals are associated with development, as well as physiological and pathological behaviors. Several conserved mitochondrial genes exist between plants and higher eukaryotes. Yet, the similarities in mitochondrial function between plant and animal species is poorly understood. Here, we show that FMT (FRIENDLY MITOCHONDRIA) from Arabidopsis thaliana, a highly conserved homolog of the mammalian CLUH (CLUSTERED MITOCHONDRIA) gene family encoding mitochondrial proteins associated with developmental alterations and adult physiological and pathological behaviors, affects whole plant morphology and development under both stressed and normal growth conditions. FMT was found to regulate mitochondrial morphology and dynamics, germination, and flowering time. It also affects leaf expansion growth, salt stress responses and hyponastic behavior, including changes in speed of hyponastic movements. Strikingly, Cluh(+/-) heterozygous knockout mice also displayed altered locomotive movements, traveling for shorter distances and had slower average and maximum speeds in the open field test. These observations indicate that homologous mitochondrial genes may play similar roles and affect homologous functions in both plants and animals. KW - Arabidopsis thaliana KW - Mitochondria KW - FMT KW - Hyponasty KW - Mice KW - CLUH; KW - Locomotion Y1 - 2022 U6 - https://doi.org/10.1007/s00018-022-04382-3 SN - 1420-682X SN - 1420-9071 VL - 79 IS - 6 PB - Springer International Publishing AG CY - Cham (ZG) ER - TY - GEN A1 - Fichtner, Franziska A1 - Barbier, Francois F. A1 - Annunziata, Maria Grazia A1 - Feil, Regina A1 - Olas, Justyna Jadwiga A1 - Müller-Röber, Bernd A1 - Stitt, Mark A1 - Beveridge, Christine A. A1 - Lunn, John Edward T1 - Regulation of shoot branching in arabidopsis by trehalose 6-phosphate T2 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe N2 - Trehalose 6-phosphate (Tre6P) is a sucrose signalling metabolite that has been implicated in regulation of shoot branching, but its precise role is not understood. We expressed tagged forms of TREHALOSE-6-PHOSPHATE SYNTHASE1 (TPS1) to determine where Tre6P is synthesized in arabidopsis (Arabidopsis thaliana), and investigated the impact of localized changes in Tre6P levels, in axillary buds or vascular tissues, on shoot branching in wild-type and branching mutant backgrounds. TPS1 is expressed in axillary buds and the subtending vasculature, as well as in the leaf and stem vasculature. Expression of a heterologous Tre6P phosphatase (TPP) to lower Tre6P in axillary buds strongly delayed bud outgrowth in long days and inhibited branching in short days. TPP expression in the vasculature also delayed lateral bud outgrowth and decreased branching. Increased Tre6P in the vasculature enhanced branching and was accompanied by higher expression of FLOWERING LOCUS T (FT) and upregulation of sucrose transporters. Increased vascular Tre6P levels enhanced branching in branched1 but not in ft mutant backgrounds. These results provide direct genetic evidence of a local role for Tre6P in regulation of axillary bud outgrowth within the buds themselves, and also connect Tre6P with systemic regulation of shoot branching via FT. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 1383 KW - Arabidopsis thaliana (arabidopsis) KW - axillary bud KW - branching KW - sucrose KW - sugar signalling KW - trehalose 6‐ phosphate (Tre6P) Y1 - 2020 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-569564 SN - 1866-8372 IS - 4 ER - TY - JOUR A1 - John, Sheeba A1 - Olas, Justyna Jadwiga A1 - Müller-Röber, Bernd T1 - Regulation of alternative splicing in response to temperature variation in plants JF - Journal of experimental botany N2 - Plants have evolved numerous molecular strategies to cope with perturbations in environmental temperature, and to adjust growth and physiology to limit the negative effects of extreme temperature. One of the strategies involves alternative splicing of primary transcripts to encode alternative protein products or transcript variants destined for degradation by nonsense-mediated decay. Here, we review how changes in environmental temperature-cold, heat, and moderate alterations in temperature-affect alternative splicing in plants, including crops. We present examples of the mode of action of various temperature-induced splice variants and discuss how these alternative splicing events enable favourable plant responses to altered temperatures. Finally, we point out unanswered questions that should be addressed to fully utilize the endogenous mechanisms in plants to adjust their growth to environmental temperature. We also indicate how this knowledge might be used to enhance crop productivity in the future. KW - alternative splicing KW - ambient temperature KW - cold KW - heat KW - plants KW - stress KW - adaptation Y1 - 2021 U6 - https://doi.org/10.1093/jxb/erab232 SN - 0022-0957 SN - 1460-2431 VL - 72 IS - 18 SP - 6150 EP - 6163 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Faisal, Muhammad B. A1 - Gechev, Tsanko S. A1 - Müller-Röber, Bernd A1 - Dijkwel, Paul P. T1 - Putative alternative translation start site-encoding nucleotides of CPR5 regulate growth and resistance JF - BMC plant biology N2 - Background The Arabidopsis CONSTITUTIVE EXPRESSER of PATHOGENESIS-RELATED GENES 5 (CPR5) has recently been shown to play a role in gating as part of the nuclear pore complex (NPC). Mutations in CPR5 cause multiple defects, including aberrant trichomes, reduced ploidy levels, reduced growth and enhanced resistance to bacterial and fungal pathogens. The pleiotropic nature of cpr5 mutations implicates that the CPR5 protein affects multiple pathways. However, little is known about the structural features that allow CPR5 to affect the different pathways. Results Our in silico studies suggest that in addition to three clusters of putative nuclear localization signals and four or five transmembrane domains, CPR5 contains two putative alternative translation start sites. To test the role of the methionine-encoding nucleotides implicated in those sites, metCPR5 cDNAs, in which the relevant nucleotides were changed to encode glutamine, were fused to the CPR5 native promoter and the constructs transformed to cpr5-2 plants to complement cpr5-compromised phenotypes. The control and metCPR5 constructs were able to complement all cpr5 phenotypes, although the extent of complementation depended on the specific complementing plant lines. Remarkably, plants transformed with metCPR5 constructs showed larger leaves and displayed reduced resistance when challenged to Pseudomonas syringae pv Pst DC3000, as compared to control plants. Thus, the methionine-encoding nucleotides regulate growth and resistance. We propose that structural features of the CPR5 N-terminus are implicated in selective gating of proteins involved in regulating the balance between growth and resistance. Conclusion Plants need to carefully balance the amount of resources used for growth and resistance. The Arabidopsis CPR5 protein regulates plant growth and immunity. Here we show that N-terminal features of CPR5 are involved in the regulation of the balance between growth and resistance. These findings may benefit efforts to improve plant yield, while maintaining optimal levels of disease resistance. KW - CPR5 KW - plant growth KW - disease resistance KW - cell death KW - arabidopsis thaliana KW - endoreduplication Y1 - 2020 U6 - https://doi.org/10.1186/s12870-020-02485-2 SN - 1471-2229 VL - 20 IS - 1 PB - BMC CY - London ER - TY - JOUR A1 - Moreno Curtidor, Catalina A1 - Annunziata, Maria Grazia A1 - Gupta, Saurabh A1 - Apelt, Federico A1 - Richard, Sarah Isabel A1 - Kragler, Friedrich A1 - Müller-Röber, Bernd A1 - Olas, Justyna Jadwiga T1 - Physiological profiling of embryos and dormant seeds in two Arabidopsis accessions reveals a metabolic switch in carbon reserve accumulation JF - Frontiers in plant science N2 - In flowering plants, sugars act as carbon sources providing energy for developing embryos and seeds. Although most studies focus on carbon metabolism in whole seeds, knowledge about how particular sugars contribute to the developmental transitions during embryogenesis is scarce. To develop a quantitative understanding of how carbon composition changes during embryo development, and to determine how sugar status contributes to final seed or embryo size, we performed metabolic profiling of hand-dissected embryos at late torpedo and mature stages, and dormant seeds, in two Arabidopsis thaliana accessions with medium [Columbia-0 (Col-0)] and large [Burren-0 (Bur-0)] seed sizes, respectively. Our results show that, in both accessions, metabolite profiles of embryos largely differ from those of dormant seeds. We found that developmental transitions from torpedo to mature embryos, and further to dormant seeds, are associated with major metabolic switches in carbon reserve accumulation. While glucose, sucrose, and starch predominantly accumulated during seed dormancy, fructose levels were strongly elevated in mature embryos. Interestingly, Bur-0 seeds contain larger mature embryos than Col-0 seeds. Fructose and starch were accumulated to significantly higher levels in mature Bur-0 than Col-0 embryos, suggesting that they contribute to the enlarged mature Bur-0 embryos. Furthermore, we found that Bur-0 embryos accumulated a higher level of sucrose compared to hexose sugars and that changes in sucrose metabolism are mediated by sucrose synthase (SUS), with SUS genes acting non-redundantly, and in a tissue-specific manner to utilize sucrose during late embryogenesis. KW - carbon KW - embryo development KW - hexoses KW - metabolites KW - sucrose KW - synthase Y1 - 2020 U6 - https://doi.org/10.3389/fpls.2020.588433 SN - 1664-462X VL - 11 PB - Frontiers Media CY - Lausanne ER - TY - JOUR A1 - Fichtner, Franziska A1 - Barbier, Francois F. A1 - Annunziata, Maria Grazia A1 - Feil, Regina A1 - Olas, Justyna Jadwiga A1 - Müller-Röber, Bernd A1 - Stitt, Mark A1 - Beveridge, Christine A. A1 - Lunn, John Edward T1 - Regulation of shoot branching in arabidopsis by trehalose 6-phosphate JF - New phytologist : international journal of plant science N2 - Trehalose 6-phosphate (Tre6P) is a sucrose signalling metabolite that has been implicated in regulation of shoot branching, but its precise role is not understood. We expressed tagged forms of TREHALOSE-6-PHOSPHATE SYNTHASE1 (TPS1) to determine where Tre6P is synthesized in arabidopsis (Arabidopsis thaliana), and investigated the impact of localized changes in Tre6P levels, in axillary buds or vascular tissues, on shoot branching in wild-type and branching mutant backgrounds. TPS1 is expressed in axillary buds and the subtending vasculature, as well as in the leaf and stem vasculature. Expression of a heterologous Tre6P phosphatase (TPP) to lower Tre6P in axillary buds strongly delayed bud outgrowth in long days and inhibited branching in short days. TPP expression in the vasculature also delayed lateral bud outgrowth and decreased branching. Increased Tre6P in the vasculature enhanced branching and was accompanied by higher expression of FLOWERING LOCUS T (FT) and upregulation of sucrose transporters. Increased vascular Tre6P levels enhanced branching in branched1 but not in ft mutant backgrounds. These results provide direct genetic evidence of a local role for Tre6P in regulation of axillary bud outgrowth within the buds themselves, and also connect Tre6P with systemic regulation of shoot branching via FT. KW - Arabidopsis thaliana (arabidopsis) KW - axillary bud KW - branching KW - sucrose KW - sugar signalling KW - trehalose 6‐ phosphate (Tre6P) Y1 - 2020 U6 - https://doi.org/10.1111/nph.17006 SN - 0028-646X SN - 1469-8137 VL - 229 IS - 4 SP - 2135 EP - 2151 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Hasnat, Muhammad Abrar A1 - Zupok, Arkadiusz A1 - Olas-Apelt, Justyna Jadwiga A1 - Müller-Röber, Bernd A1 - Leimkühler, Silke T1 - A-type carrier proteins are involved in [4Fe-4S] cluster insertion into the radical S-adenosylmethionine protein MoaA for the synthesis of active molybdoenzymes JF - Journal of bacteriology N2 - Iron sulfur (Fe-S) clusters are important biological cofactors present in proteins with crucial biological functions, from photosynthesis to DNA repair, gene expression, and bioenergetic processes. For the insertion of Fe-S clusters into proteins, A-type carrier proteins have been identified. So far, three of them have been characterized in detail in Escherichia coli, namely, IscA, SufA, and ErpA, which were shown to partially replace each other in their roles in [4Fe-4S] cluster insertion into specific target proteins. To further expand the knowledge of [4Fe-4S] cluster insertion into proteins, we analyzed the complex Fe-S cluster-dependent network for the synthesis of the molybdenum cofactor (Moco) and the expression of genes encoding nitrate reductase in E. coli. Our studies include the identification of the A-type carrier proteins ErpA and IscA, involved in [4Fe-4S] cluster insertion into the radical Sadenosyl-methionine (SAM) enzyme MoaA. We show that ErpA and IscA can partially replace each other in their role to provide [4Fe-4S] clusters for MoaA. Since most genes expressing molybdoenzymes are regulated by the transcriptional regulator for fumarate and nitrate reduction (FNR) under anaerobic conditions, we also identified the proteins that are crucial to obtain an active FNR under conditions of nitrate respiration. We show that ErpA is essential for the FNR-dependent expression of the narGHJI operon, a role that cannot be compensated by IscA under the growth conditions tested. SufA does not appear to have a role in Fe-S cluster insertion into MoaA or FNR under anaerobic growth employing nitrate respiration, based on the low level of gene expression.
IMPORTANCE Understanding the assembly of iron-sulfur (Fe-S) proteins is relevant to many fields, including nitrogen fixation, photosynthesis, bioenergetics, and gene regulation. Remaining critical gaps in our knowledge include how Fe-S clusters are transferred to their target proteins and how the specificity in this process is achieved, since different forms of Fe-S clusters need to be delivered to structurally highly diverse target proteins. Numerous Fe-S carrier proteins have been identified in prokaryotes like Escherichia coli, including ErpA, IscA, SufA, and NfuA. In addition, the diverse Fe-S cluster delivery proteins and their target proteins underlie a complex regulatory network of expression, to ensure that both proteins are synthesized under particular growth conditions. KW - iron-sulfur clusters KW - Moco biosynthesis KW - MoaA KW - A-type carrier protein KW - FNR KW - nitrate reductase KW - molybdenum cofactor Y1 - 2021 U6 - https://doi.org/10.1128/JB.00086-21 SN - 1098-5530 VL - 203 IS - 12 PB - American Society for Microbiology CY - Washington ER - TY - JOUR A1 - Shubchynskyy, Volodymyr A1 - Boniecka, Justyna A1 - Schweighofer, Alois A1 - Simulis, Justinas A1 - Kvederaviciute, Kotryna A1 - Stumpe, Michael A1 - Mauch, Felix A1 - Balazadeh, Salma A1 - Müller-Röber, Bernd A1 - Boutrot, Freddy A1 - Zipfel, Cyril A1 - Meskiene, Irute T1 - Protein phosphatase AP2C1 negatively regulates basal resistance and defense responses to Pseudomonas syringae JF - Journal of experimental botany N2 - Mitogen-activated protein kinases (MAPKs) mediate plant immune responses to pathogenic bacteria. However, less is known about the cell autonomous negative regulatory mechanism controlling basal plant immunity. We report the biological role of Arabidopsis thaliana MAPK phosphatase AP2C1 as a negative regulator of plant basal resistance and defense responses to Pseudomonas syringae. AP2C2, a closely related MAPK phosphatase, also negatively controls plant resistance. Loss of AP2C1 leads to enhanced pathogen-induced MAPK activities, increased callose deposition in response to pathogen-associated molecular patterns or to P. syringae pv. tomato (Pto) DC3000, and enhanced resistance to bacterial infection with Pto. We also reveal the impact of AP2C1 on the global transcriptional reprogramming of transcription factors during Pto infection. Importantly, ap2c1 plants show salicylic acid-independent transcriptional reprogramming of several defense genes and enhanced ethylene production in response to Pto. This study pinpoints the specificity of MAPK regulation by the different MAPK phosphatases AP2C1 and MKP1, which control the same MAPK substrates, nevertheless leading to different downstream events. We suggest that precise and specific control of defined MAPKs by MAPK phosphatases during plant challenge with pathogenic bacteria can strongly influence plant resistance. KW - Callose KW - defense genes KW - MAPK KW - MAPK phosphatase KW - PAMP KW - PP2C phosphatase KW - Pseudomonas syringae KW - salicylic acid KW - transcription factors Y1 - 2017 U6 - https://doi.org/10.1093/jxb/erw485 SN - 0022-0957 SN - 1460-2431 VL - 68 IS - 5 SP - 1169 EP - 1183 PB - Oxford Univ. Press CY - Oxford ER - TY - JOUR A1 - Sharma, Niharika A1 - Dang, Trang Minh A1 - Singh, Namrata A1 - Ruzicic, Slobodan A1 - Müller-Röber, Bernd A1 - Baumann, Ute A1 - Heuer, Sigrid T1 - Allelic variants of OsSUB1A cause differential expression of transcription factor genes in response to submergence in rice JF - Rice N2 - Background: Flooding during seasonal monsoons affects millions of hectares of rice-cultivated areas across Asia. Submerged rice plants die within a week due to lack of oxygen, light and excessive elongation growth to escape the water. Submergence tolerance was first reported in an aus-type rice landrace, FR13A, and the ethylene-responsive transcription factor (TF) gene SUB1A-1 was identified as the major tolerance gene. Intolerant rice varieties generally lack the SUB1A gene but some intermediate tolerant varieties, such as IR64, carry the allelic variant SUB1A-2. Differential effects of the two alleles have so far not been addressed. As a first step, we have therefore quantified and compared the expression of nearly 2500 rice TF genes between IR64 and its derived tolerant near isogenic line IR64-Sub1, which carries the SUB1A-1 allele. Gene expression was studied in internodes, where the main difference in expression between the two alleles was previously shown. Results: Nineteen and twenty-six TF genes were identified that responded to submergence in IR64 and IR64-Sub1, respectively. Only one gene was found to be submergence-responsive in both, suggesting different regulatory pathways under submergence in the two genotypes. These differentially expressed genes (DEGs) mainly included MYB, NAC, TIFY and Zn-finger TFs, and most genes were downregulated upon submergence. In IR64, but not in IR64-Sub1, SUB1B and SUB1C, which are also present in the Sub1 locus, were identified as submergence responsive. Four TFs were not submergence responsive but exhibited constitutive, genotype-specific differential expression. Most of the identified submergence responsive DEGs are associated with regulatory hormonal pathways, i.e. gibberellins (GA), abscisic acid (ABA), and jasmonic acid (JA), apart from ethylene. An in-silico promoter analysis of the two genotypes revealed the presence of allele-specific single nucleotide polymorphisms, giving rise to ABRE, DRE/CRT, CARE and Site II cis-elements, which can partly explain the observed differential TF gene expression. Conclusion: This study identified new gene targets with the potential to further enhance submergence tolerance in rice and provides insights into novel aspects of SUB1A-mediated tolerance. KW - Submergence tolerance KW - SUB1A KW - Rice KW - Transcription factors Y1 - 2018 U6 - https://doi.org/10.1186/s12284-017-0192-z SN - 1939-8425 SN - 1939-8433 VL - 11 IS - 2 PB - Springer Open CY - London ER - TY - JOUR A1 - Durgud, Meriem A1 - Gupta, Saurabh A1 - Ivanov, Ivan A1 - Omidbakhshfard, Mohammad Amin A1 - Benina, Maria A1 - Alseekh, Saleh A1 - Staykov, Nikola A1 - Hauenstein, Mareike A1 - Dijkwel, Paul P. A1 - Hortensteiner, Stefan A1 - Toneva, Valentina A1 - Brotman, Yariv A1 - Fernie, Alisdair R. A1 - Müller-Röber, Bernd A1 - Gechev, Tsanko S. T1 - Molecular Mechanisms Preventing Senescence in Response to Prolonged Darkness in a Desiccation-Tolerant Plant JF - Plant physiology : an international journal devoted to physiology, biochemistry, cellular and molecular biology, biophysics and environmental biology of plants N2 - The desiccation-tolerant plant Haberlea rhodopensis can withstand months of darkness without any visible senescence. Here, we investigated the molecular mechanisms of this adaptation to prolonged (30 d) darkness and subsequent return to light. H. rhodopensis plants remained green and viable throughout the dark treatment. Transcriptomic analysis revealed that darkness regulated several transcription factor (TF) genes. Stress-and autophagy-related TFs such as ERF8, HSFA2b, RD26, TGA1, and WRKY33 were up-regulated, while chloroplast-and flowering-related TFs such as ATH1, COL2, COL4, RL1, and PTAC7 were repressed. PHYTOCHROME INTERACTING FACTOR4, a negative regulator of photomorphogenesis and promoter of senescence, also was down-regulated. In response to darkness, most of the photosynthesis-and photorespiratory-related genes were strongly down-regulated, while genes related to autophagy were up-regulated. This occurred concomitant with the induction of SUCROSE NON-FERMENTING1-RELATED PROTEIN KINASES (SnRK1) signaling pathway genes, which regulate responses to stress-induced starvation and autophagy. Most of the genes associated with chlorophyll catabolism, which are induced by darkness in dark-senescing species, were either unregulated (PHEOPHORBIDE A OXYGENASE, PAO; RED CHLOROPHYLL CATABOLITE REDUCTASE, RCCR) or repressed (STAY GREEN-LIKE, PHEOPHYTINASE, and NON-YELLOW COLORING1). Metabolite profiling revealed increases in the levels of many amino acids in darkness, suggesting increased protein degradation. In darkness, levels of the chloroplastic lipids digalactosyldiacylglycerol, monogalactosyldiacylglycerol, phosphatidylglycerol, and sulfoquinovosyldiacylglycerol decreased, while those of storage triacylglycerols increased, suggesting degradation of chloroplast membrane lipids and their conversion to triacylglycerols for use as energy and carbon sources. Collectively, these data show a coordinated response to darkness, including repression of photosynthetic, photorespiratory, flowering, and chlorophyll catabolic genes, induction of autophagy and SnRK1 pathways, and metabolic reconfigurations that enable survival under prolonged darkness. Y1 - 2018 U6 - https://doi.org/10.1104/pp.18.00055 SN - 0032-0889 SN - 1532-2548 VL - 177 IS - 3 SP - 1319 EP - 1338 PB - American Society of Plant Physiologists CY - Rockville ER - TY - JOUR A1 - Hochrein, Lena A1 - Mitchell, Leslie A. A1 - Schulz, Karina A1 - Messerschmidt, Katrin A1 - Müller-Röber, Bernd T1 - L-SCRaMbLE as a tool for light-controlled Cre-mediated recombination in yeast JF - Nature Communications N2 - The synthetic yeast genome constructed by the International Synthetic Yeast Sc2.0 consortium adds thousands of loxPsym recombination sites to all 16 redesigned chromosomes, allowing the shuffling of Sc2.0 chromosome parts by the Cre-loxP recombination system thereby enabling genome evolution experiments. Here, we present L-SCRaMbLE, a lightcontrolled Cre recombinase for use in the yeast Saccharomyces cerevisiae. L-SCRaMbLE allows tight regulation of recombinase activity with up to 179-fold induction upon exposure to red light. The extent of recombination depends on induction time and concentration of the chromophore phycocyanobilin (PCB), which can be easily adjusted. The tool presented here provides improved recombination control over the previously reported estradiol-dependent SCRaMbLE induction system, mediating a larger variety of possible recombination events in SCRaMbLE-ing a reporter plasmid. Thereby, L-SCRaMbLE boosts the potential for further customization and provides a facile application for use in the S. cerevisiae genome reengineering project Sc2.0 or in other recombination-based systems. Y1 - 2018 U6 - https://doi.org/10.1038/s41467-017-02208-6 SN - 2041-1723 VL - 9 PB - Nature Publ. Group CY - London ER - TY - GEN A1 - Balazadeh, Salma A1 - Müller-Röber, Bernd T1 - A balance to death T2 - Nature plants N2 - Leaf senescence plays a crucial role in nutrient recovery in late-stage plant development and requires vast transcriptional reprogramming by transcription factors such as ORESARA1 (ORE1). A proteolytic mechanism is now found to control ORE1 degradation, and thus senescence, during nitrogen starvation. Y1 - 2018 U6 - https://doi.org/10.1038/s41477-018-0279-6 SN - 2055-026X SN - 2055-0278 VL - 4 IS - 11 SP - 863 EP - 864 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Naseri, Gita A1 - Behrend, Jessica A1 - Rieper, Lisa A1 - Müller-Röber, Bernd T1 - COMPASS for rapid combinatorial optimization of biochemical pathways based on artificial transcription factors JF - Nature Communications N2 - Balanced expression of multiple genes is central for establishing new biosynthetic pathways or multiprotein cellular complexes. Methods for efficient combinatorial assembly of regulatory sequences (promoters) and protein coding sequences are therefore highly wanted. Here, we report a high-throughput cloning method, called COMPASS for COMbinatorial Pathway ASSembly, for the balanced expression of multiple genes in Saccharomyces cerevisiae. COMPASS employs orthogonal, plant-derived artificial transcription factors (ATFs) and homologous recombination-based cloning for the generation of thousands of individual DNA constructs in parallel. The method relies on a positive selection of correctly assembled pathway variants from both, in vivo and in vitro cloning procedures. To decrease the turnaround time in genomic engineering, COMPASS is equipped with multi-locus CRISPR/Cas9-mediated modification capacity. We demonstrate the application of COMPASS by generating cell libraries producing n-carotene and co-producing p-ionone and biosensor-responsive naringenin. COMPASS will have many applications in synthetic biology projects that require gene expression balancing. Y1 - 2019 U6 - https://doi.org/10.1038/s41467-019-10224-x SN - 2041-1723 VL - 10 PB - Nature Publ. Group CY - London ER - TY - GEN A1 - Das Gupta, Mainak A1 - Roesch, Florian A1 - Hochrein, Lena A1 - Machens, Fabian A1 - Müller-Röber, Bernd T1 - Facilitating Genome Engineering Through RNP-mediated Precise Gene Targeting T2 - In Vitro Cellular & Developmental Biology - Plant Y1 - 2019 SN - 1054-5476 SN - 1475-2689 VL - 55 IS - 4 SP - 481 EP - 481 PB - Springer CY - New York ER - TY - JOUR A1 - Yang, Lei A1 - Perrera, Valentina A1 - Saplaoura, Eleftheria A1 - Apelt, Federico A1 - Bahin, Mathieu A1 - Kramdi, Amira A1 - Olas, Justyna Jadwiga A1 - Müller-Röber, Bernd A1 - Sokolowska, Ewelina A1 - Zhang, Wenna A1 - Li, Runsheng A1 - Pitzalis, Nicolas A1 - Heinlein, Manfred A1 - Zhang, Shoudong A1 - Genovesio, Auguste A1 - Colot, Vincent A1 - Kragler, Friedrich T1 - m(5)C Methylation Guides Systemic Transport of Messenger RNA over Graft Junctions in Plants JF - Current biology N2 - In plants, transcripts move to distant body parts to potentially act as systemic signals regulating development and growth. Thousands of messenger RNAs (mRNAs) are transported across graft junctions via the phloem to distinct plant parts. Little is known regarding features, structural motifs, and potential base modifications of transported transcripts and how these may affect their mobility. We identified Arabidopsis thalianam RNAs harboring the modified base 5-methylcytosine (m(5)C) and found that these are significantly enriched in mRNAs previously described as mobile, moving over graft junctions to distinct plant parts. We confirm this finding with graft-mobile methylated mRNAs TRANSLATIONALLY CONTROLLED TUMOR PROTEIN 1 (TCTP1) and HEAT SHOCK COGNATE PROTEIN 70.1 (HSC70.1), whose mRNA transport is diminished in mutants deficient in m(5)C mRNA methylation. Together, our results point toward an essential role of cytosine methylation in systemic mRNA mobility in plants and that TCTP1 mRNA mobility is required for its signaling function. Y1 - 2019 U6 - https://doi.org/10.1016/j.cub.2019.06.042 SN - 0960-9822 SN - 1879-0445 VL - 29 IS - 15 SP - 2465 EP - 2476.e5 PB - Cell Press CY - Cambridge ER - TY - JOUR A1 - Dong, Yanni A1 - Gupta, Saurabh A1 - Sievers, Rixta A1 - Wargent, Jason J. A1 - Wheeler, David A1 - Putterill, Joanna A1 - Macknight, Richard A1 - Gechev, Tsanko S. A1 - Müller-Röber, Bernd A1 - Dijkwel, Paul P. T1 - Genome draft of the Arabidopsis relative Pachycladon cheesemanii reveals environment JF - BMC genomics N2 - BackgroundPachycladon cheesemanii is a close relative of Arabidopsis thaliana and is an allotetraploid perennial herb which is widespread in the South Island of New Zealand. It grows at altitudes of up to 1000m where it is subject to relatively high levels of ultraviolet (UV)-B radiation. To gain first insights into how Pachycladon copes with UV-B stress, we sequenced its genome and compared the UV-B tolerance of two Pachycladon accessions with those of two A. thaliana accessions from different altitudes.ResultsA high-quality draft genome of P. cheesemanii was assembled with a high percentage of conserved single-copy plant orthologs. Synteny analysis with genomes from other species of the Brassicaceae family found a close phylogenetic relationship of P. cheesemanii with Boechera stricta from Brassicaceae lineage I. While UV-B radiation caused a greater growth reduction in the A. thaliana accessions than in the P. cheesemanii accessions, growth was not reduced in one P. cheesemanii accession. The homologues of A. thaliana UV-B radiation response genes were duplicated in P. cheesemanii, and an expression analysis of those genes indicated that the tolerance mechanism in P. cheesemanii appears to differ from that in A. thaliana.ConclusionAlthough the P. cheesemanii genome shows close similarity with that of A. thaliana, it appears to have evolved novel strategies allowing the plant to tolerate relatively high UV-B radiation. KW - Abiotic stress KW - Arabidopsis KW - Genome assembly KW - Pachycladon KW - UV-B tolerance Y1 - 2019 U6 - https://doi.org/10.1186/s12864-019-6084-4 SN - 1471-2164 VL - 20 IS - 1 PB - BMC CY - London ER - TY - JOUR A1 - Naseri, Gita A1 - Balazadeh, Salma A1 - Machens, Fabian A1 - Kamranfar, Iman A1 - Messerschmidt, Katrin A1 - Müller-Röber, Bernd T1 - Plant-Derived Transcription Factors for Orthologous Regulation of Gene Expression in the Yeast Saccharomyces cerevisiae JF - ACS synthetic biology N2 - Control of gene expression by transcription factors (TFs) is central in many synthetic biology projects for which a tailored expression of one or multiple genes is often needed. As TFs from evolutionary distant organisms are unlikely to affect gene expression in a host of choice, they represent excellent candidates for establishing orthogonal control systems. To establish orthogonal regulators for use in yeast (Saccharomyces cerevisiae), we chose TFs from the plant Arabidopsis thaliana. We established a library of 106 different combinations of chromosomally integrated TFs, activation domains (yeast GAL4 AD, herpes simplex virus VP64, and plant EDLL) and synthetic promoters harboring cognate cis regulatory motifs driving a yEGFP reporter. Transcriptional output of the different driver/reporter combinations varied over a wide spectrum, with EDLL being a considerably stronger transcription activation domain in yeast than the GAL4 activation domain, in particular when fused to Arabidopsis NAC TFs. Notably, the strength of several NAC-EDLL fusions exceeded that of the strong yeast TDH3 promoter by 6- to 10-fold. We furthermore show that plant TFs can be used to build regulatory systems encoded by centromeric or episomal plasmids. Our library of TF-DNA binding site combinations offers an excellent tool for diverse synthetic biology applications in yeast. KW - Arabidopsis thaliana KW - artificial transcription factor KW - NAC transcription factor KW - synthetic biology KW - plant Y1 - 2017 U6 - https://doi.org/10.1021/acssynbio.7b00094 SN - 2161-5063 VL - 6 SP - 1742 EP - 1756 PB - American Chemical Society CY - Washington ER - TY - JOUR A1 - Czarnocka, Weronika A1 - Van Der Kelen, Katrien A1 - Willems, Patrick A1 - Szechynska-Hebda, Magdalena A1 - Shahnejat-Bushehri, Sara A1 - Balazadeh, Salma A1 - Rusaczonek, Anna A1 - Müller-Röber, Bernd A1 - Van Breusegem, Frank A1 - Karpinski, Stanislaw T1 - The dual role of LESION SIMULATING DISEASE 1 as a condition-dependent scaffold protein and transcription regulator JF - Plant, cell & environment : cell physiology, whole-plant physiology, community physiology N2 - Since its discovery over two decades ago as an important cell death regulator in Arabidopsis thaliana, the role of LESION SIMULATING DISEASE 1 (LSD1) has been studied intensively within both biotic and abiotic stress responses as well as with respect to plant fitness regulation. However, its molecular mode of action remains enigmatic. Here, we demonstrate that nucleo-cytoplasmic LSD1 interacts with a broad range of other proteins that are engaged in various molecular pathways such as ubiquitination, methylation, cell cycle control, gametogenesis, embryo development and cell wall formation. The interaction of LSD1 with these partners is dependent on redox status, as oxidative stress significantly changes the quantity and types of LSD1-formed complexes. Furthermore, we show that LSD1 regulates the number and size of leaf mesophyll cells and affects plant vegetative growth. Importantly, we also reveal that in addition to its function as a scaffold protein, LSD1 acts as a transcriptional regulator. Taken together, our results demonstrate that LSD1 plays a dual role within the cell by acting as a condition-dependent scaffold protein and as a transcription regulator. KW - Arabidopsis KW - thaliana KW - dry weight KW - LSD1 KW - oxidative stress KW - protein interaction KW - transcription regulation Y1 - 2017 U6 - https://doi.org/10.1111/pce.12994 SN - 0140-7791 SN - 1365-3040 VL - 40 SP - 2644 EP - 2662 PB - Wiley CY - Hoboken ER - TY - JOUR A1 - Omranian, Nooshin A1 - Eloundou-Mbebi, Jeanne Marie Onana A1 - Müller-Röber, Bernd A1 - Nikoloski, Zoran T1 - Gene regulatory network inference using fused LASSO on multiple data sets JF - Scientific reports N2 - Devising computational methods to accurately reconstruct gene regulatory networks given gene expression data is key to systems biology applications. Here we propose a method for reconstructing gene regulatory networks by simultaneous consideration of data sets from different perturbation experiments and corresponding controls. The method imposes three biologically meaningful constraints: (1) expression levels of each gene should be explained by the expression levels of a small number of transcription factor coding genes, (2) networks inferred from different data sets should be similar with respect to the type and number of regulatory interactions, and (3) relationships between genes which exhibit similar differential behavior over the considered perturbations should be favored. We demonstrate that these constraints can be transformed in a fused LASSO formulation for the proposed method. The comparative analysis on transcriptomics time-series data from prokaryotic species, Escherichia coli and Mycobacterium tuberculosis, as well as a eukaryotic species, mouse, demonstrated that the proposed method has the advantages of the most recent approaches for regulatory network inference, while obtaining better performance and assigning higher scores to the true regulatory links. The study indicates that the combination of sparse regression techniques with other biologically meaningful constraints is a promising framework for gene regulatory network reconstructions. Y1 - 2016 U6 - https://doi.org/10.1038/srep20533 SN - 2045-2322 VL - 6 PB - Nature Publ. Group CY - London ER - TY - JOUR A1 - Sedaghatmehr, Mastoureh A1 - Müller-Röber, Bernd A1 - Balazadeh, Salma T1 - The plastid metalloprotease FtsH6 and small heat shock protein HSP21 jointly regulate thermomemory in Arabidopsis JF - Nature Communications N2 - Acquired tolerance to heat stress is an increased resistance to elevated temperature following a prior exposure to heat. The maintenance of acquired thermotolerance in the absence of intervening stress is called ‘thermomemory’ but the mechanistic basis for this memory is not well defined. Here we show that Arabidopsis HSP21, a plastidial small heat shock protein that rapidly accumulates after heat stress and remains abundant during the thermomemory phase, is a crucial component of thermomemory. Sustained memory requires that HSP21 levels remain high. Through pharmacological interrogation and transcriptome profiling, we show that the plastid-localized metalloprotease FtsH6 regulates HSP21 abundance. Lack of a functional FtsH6 protein promotes HSP21 accumulation during the later stages of thermomemory and increases thermomemory capacity. Our results thus reveal the presence of a plastidial FtsH6–HSP21 control module for thermomemory in plants. Y1 - 2016 U6 - https://doi.org/10.1038/ncomms12439 SN - 2041-1723 VL - 7 PB - Nature Publ. Group CY - London ER -