scholarly journals Origins and Immunity Networking Functions of EDS1 Family Proteins

2020 ◽  
Vol 58 (1) ◽  
pp. 253-276 ◽  
Author(s):  
Dmitry Lapin ◽  
Deepak D. Bhandari ◽  
Jane E. Parker

The EDS1 family of structurally unique lipase-like proteins EDS1, SAG101, and PAD4 evolved in seed plants, on top of existing phytohormone and nucleotide-binding–leucine-rich-repeat (NLR) networks, to regulate immunity pathways against host-adapted biotrophic pathogens. Exclusive heterodimers between EDS1 and SAG101 or PAD4 create essential surfaces for resistance signaling. Phylogenomic information, together with functional studies in Arabidopsis and tobacco, identify a coevolved module between the EDS1–SAG101 heterodimer and coiled-coil (CC) HET-S and LOP-B (CCHELO) domain helper NLRs that is recruited by intracellular Toll-interleukin1-receptor (TIR) domain NLR receptors to confer host cell death and pathogen immunity. EDS1–PAD4 heterodimers have a different and broader activity in basal immunity that transcriptionally reinforces local and systemic defenses triggered by various NLRs. Here, we consider EDS1 family protein functions across seed plant lineages in the context of networking with receptor and helper NLRs and downstream resistance machineries. The different modes of action and pathway connectivities of EDS1 family members go some way to explaining their central role in biotic stress resilience.

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 852
Author(s):  
Viacheslav V. Senichkin ◽  
Evgeniia A. Prokhorova ◽  
Boris Zhivotovsky ◽  
Gelina S. Kopeina

Subcellular fractionation approaches remain an indispensable tool among a large number of biochemical methods to facilitate the study of specific intracellular events and characterization of protein functions. During apoptosis, the best-known form of programmed cell death, numerous proteins are translocated into and from the nucleus. Therefore, suitable biochemical techniques for the subcellular fractionation of apoptotic cells are required. However, apoptotic bodies and cell fragments might contaminate the fractions upon using the standard protocols. Here, we compared different nucleus/cytoplasm fractionation methods and selected the best-suited approach for the separation of nuclear and cytoplasmic contents. The described methodology is based on stepwise lysis of cells and washing of the resulting nuclei using non-ionic detergents, such as NP-40. Next, we validated this approach for fractionation of cells treated with various apoptotic stimuli. Finally, we demonstrated that nuclear fraction could be further subdivided into nucleosolic and insoluble subfractions, which is crucial for the isolation and functional studies of various proteins. Altogether, we developed a method for simple and efficient nucleus/cytoplasm fractionation of both normal and apoptotic cells.


2021 ◽  
Author(s):  
Adeline Harant ◽  
Hsuan Pai ◽  
Toshiyuki Sakai ◽  
Sophien Kamoun ◽  
Hiroaki Adachi

Abstract Nicotiana benthamiana has emerged as a complementary experimental system to Arabidopsis thaliana. It enables fast-forward in vivo analyses primarily through transient gene expression and is particularly popular in the study of plant immunity. Recently, our understanding of nucleotide-binding leucine-rich repeat (NLR) plant immune receptors has greatly advanced following the discovery of the Arabidopsis HOPZ-ACTIVATED RESISTANCE1 (ZAR1) resistosome. Here, we describe a vector system of 72 plasmids that enables functional studies of the ZAR1 resistosome in N. benthamiana. We showed that ZAR1 stands out among the coiled coil class of NLRs (CC-NLRs) for being highly conserved across distantly related dicot plant species and confirmed NbZAR1 as the N. benthamiana ortholog of Arabidopsis ZAR1. Effector-activated and autoactive NbZAR1 trigger the cell death response in N. benthamiana and this activity is dependent on a functional N-terminal α1 helix. C-terminally tagged NbZAR1 remains functional in N. benthamiana, thus enabling cell biology and biochemical studies in this plant system. We conclude that the NbZAR1 open source pZA plasmid collection forms an additional experimental system to Arabidopsis for in planta resistosome studies.


2016 ◽  
Author(s):  
Florencia Berruezo ◽  
Flavio S. J. de Souza ◽  
Pablo I. Picca ◽  
Sergio I. Nemirovsky ◽  
Leandro Martinez-Tosar ◽  
...  

AbstractMicroRNAs (miRNAs) are short, single stranded RNA molecules that regulate the stability and translation of messenger RNAs in diverse eukaryotic groups. Several miRNA genes are of ancient origin and have been maintained in the genomes of animal and plant taxa for hundreds of millions of years, and functional studies indicate that ancient miRNAs play key roles in development and physiology. In the last decade, genome and small RNA (sRNA) sequencing of several plant species have helped unveil the evolutionary history of land plant miRNAs. Land plants are divided into bryophytes (liverworts, mosses), lycopods (clubmosses and spikemosses), monilophytes (ferns and horsetails), gymnosperms (cycads, conifers and allies) and angiosperms (flowering plants). Among these, the fern group occupies a key phylogenetic position, since it represents the closest extant cousin taxon of seed plants, i.e. gymno- and angiosperms. However, in spite of their evolutionary, economic and ecological importance, no fern genome has been sequenced yet and few genomic resources are available for this group. Here, we sequenced the small RNA fraction of an epiphytic South American fern, Pleopeltis minima (Polypodiaceae), and compared it to plant miRNA databases, allowing for the identification of miRNA families that are shared by all land plants, shared by all vascular plants (tracheophytes) or shared by euphyllophytes (ferns and seed plants) only. Using the recently described transcriptome of another fern, Lygodium japonicum, we also estimated the degree of conservation of fern miRNA targets in relation to other plant groups. Our results pinpoint the origin of several miRNA families in the land plant evolutionary tree with more precision and are a resource for future genomic and functional studies of fern miRNAs.


2019 ◽  
Vol 20 (12) ◽  
pp. 2997 ◽  
Author(s):  
Xuda Ma ◽  
Yamei Dang ◽  
Xiaowen Shao ◽  
Xuechun Chen ◽  
Fei Wu ◽  
...  

Actin filaments are a major component of the cytoskeleton in eukaryotic cells and play an important role in cancer metastasis. Dynamics and reorganization of actin filaments are regulated by numerous regulators, including Rho GTPases, PAKs (p21-activated kinases), ROCKs (Rho-associated coiled-coil containing kinases), LIMKs (LIM domain kinases), and SSH1 (slingshot family protein phosphate 1). Ubiquitination, as a ubiquitous post-transcriptional modification, deceases protein levels of actin cytoskeleton regulatory factors and thereby modulates the actin cytoskeleton. There is increasing evidence showing cytoskeleton regulation by long noncoding RNAs (lncRNAs) in cancer metastasis. However, which E3 ligases are activated for the ubiquitination of actin-cytoskeleton regulators involved in tumor metastasis remains to be fully elucidated. Moreover, it is not clear how lncRNAs influence the expression of actin cytoskeleton regulators. Here, we summarize physiological and pathological mechanisms of lncRNAs and ubiquitination control mediators of actin cytoskeleton regulators which that are involved in tumorigenesis and tumor progression. Finally, we briefly discuss crosstalk between ubiquitination and lncRNA control mediators of actin-cytoskeleton regulators in cancer.


2019 ◽  
Author(s):  
Xin Zhang

AbstractIn the ovule evolution, the integument is the most attention point in discussion as a morphologic character of the seed plants. There are several theories and hypotheses about the origin of the integument were presented in the history. However, the development and function of the ovule envelopes are not so clear until now. The development of thehe basal gymnosperms Cycas and Zamia were to investigated, especially of the integument to complement the existing knowledge in seed plants. The development of ovules of seed plant is documented with morphological and anatomical using LM and SEM.The nucellar beak found in Zamia is a structure that has not been recorded previously. It protrudes from the micropyle at pollination and may be the primary acceptor for pollen. There are striking similarities to the lagenostom or salpinx in Lyginopteridatae. There may be an evolutionary way to interpret the pollination drop existing in the Lyginopteridatae. Probably the nucellar beak of Cycads, even Ginkgoales have the same function with the lagenostom or salpinx of the Lyginopteridatea. Unfortunately, pollen and transport inside the pollination chambers have not been observed. Further analysis of this unusual structure seems to be very important.


2018 ◽  
pp. 18-24
Author(s):  
V. Doronin ◽  
Yu. Kravchenko ◽  
V. Dryga ◽  
M. Budovsky ◽  
V. Doronin

In the intensive arable farming the productivity of sugar beets depends on many factors: soil-climatic conditions, introduction of highly productive hybrids, quality pre-sowing seed treatment, use of updated machinery and technologies, fertilizers, reliable plant protection, high technological processing at sugar mills and others. All the mentioned factors can reduce the productivity of sugar beets considerably, but it is not possible to reach a maximum potential of crop productivity without the use of good-quality seeds of the latest hybrids. The quality of sugar beet seeds is formed during the growth, and it depends on several factors, and first of all on the quality of mother root crops – their mass, sizes, turgor, disease damage and others. Laboratory trials were carried out at the Institute of bio-energy crops and sugar beets of Ukraine’s NAAS, field trials were carried out in the conditions of Ivanivka research-breeding station in the years of 2016–2018. The trial scheme envisaged the planting of root crops, which were not damaged with root rot and damaged with root rot, a focal point of rot being removed before planting. During vegetation, seed plants were treated with fungicides at a bud stage (one–time treatment) and at a bud stage and the beginning of flowering (two–time treatment). Fungicides were not used in the control. Mother root crops, seed plants and seeds of a sugar beet hybrid, developed on the basis of cytoplasm male sterility Romul, were the object of the research. In the field trials the following was determined: viability of the root crops (the number of the emerged crops as to the number of the panted root crops) and the seed plant density at a full emergence stage and before harvesting. Agro-biological evaluation of the seed plant condition was made before harvesting – the number of non-productive seed plants (dried, infertile ones and others), the number of disease-damaged plants and the degree of their development. A record plot area of one replication was 20 m2 in field trials, a fourfold replication. The technology of seed plant cultivation is conventional, in accordance with the recommendations of the Institute of bio-energy crops and sugar beets of Ukraine’s NAAS. The paper covered some peculiarities of the yield formation and the quality of sugar beet seeds, depending on the quality of mother root crops. It was proved that the use of the root crops, partially damaged with rot during storage, and even when a focal point of rot was removed before harvesting, had an effect on the root crop viability (which was by 4.4 % lower), the formation of more non-productive plants – by 1.6–1.8 times – due to the increase of prematurely dried plants and infertile ones, which resulted in a serious yield decease – by 0.09-0.13 t/ha and seed quality – viability by 4 % and emergence by 6 %, as compared with the variants where health root crops were planted. Two-time spraying of the seed plants with fungicides resulted in the increase of the yield capacity and the seed quality in the variants where only partially rot-damaged root crops were planted. A tendency to the increase of the mentioned indicators, both in the variants with healthy root crops planted and with partially damaged root crops planted, was recorded under one-time spraying of the seed plants with fungicides. The research of the seeds gathered from the seed plants, which were grown from the root crops, disease-damaged and healthy ones, confirmed the results of the field trials. The viability of the seeds, received from the healthy (not disease-damaged) root crops was higher by 9 %, emergence – by 5 %, as compared with the root crops, partially rot-damaged, focal points of which were removed before planting. Key words: mother root crops, rot, productive seed plants, yield capacity, viability, germination.


eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Upendo Lupanga ◽  
Rachel Röhrich ◽  
Jana Askani ◽  
Stefan Hilmer ◽  
Christiane Kiefer ◽  
...  

The V-ATPase is a versatile proton-pump found in a range of endomembrane compartments yet the mechanisms governing its differential targeting remain to be determined. In Arabidopsis, VHA-a1 targets the V-ATPase to the TGN/EE whereas VHA-a2 and VHA-a3 are localized to the tonoplast. We report here that the VHA-a1 targeting domain serves as both an ER-exit and as a TGN/EE-retention motif and is conserved among seed plants. In contrast, Marchantia encodes a single VHA-isoform that localizes to the TGN/EE and the tonoplast in Arabidopsis. Analysis of CRISPR/Cas9 generated null alleles revealed that VHA-a1 has an essential function for male gametophyte development but acts redundantly with the tonoplast isoforms during vegetative growth. We propose that in the absence of VHA-a1, VHA-a3 is partially re-routed to the TGN/EE. Our findings contribute to understanding the evolutionary origin of V-ATPase targeting and provide a striking example that differential localization does not preclude functional redundancy.


2020 ◽  
Author(s):  
Jamal Saad ◽  
Nassim Hammoudi ◽  
Rita Zgheib ◽  
Hussein Anani ◽  
Michel Drancourt

AbstractThe reason why severe cases of Buruli ulcers caused by Mycobacterium ulcerans are emerging in some South Australia counties has not been determined. In this study, we measured the diversity of M. ulcerans complex whole genome sequences (WGS) and reported a marker of this diversity. Using this marker as a probe, we compared WGS diversity in Buruli ulcer-epidemic South Australia counties versus non-epidemic Australian counties and further refined comparisons at the level of counties where severe Buruli ulcer cases have been reported. Analyzing 218 WGS (35 complete and 183 reconstructed WGS, including 174 Australian WGS) yielded 15 M. ulcerans complex genotypes, including three genotypes specific to Australia and one genotype specific to South Australia. A 1,068-bp PPE family protein gene exhibiting genotype-specific sequence variations was employed to further probe 13 minority clones hidden in sequence reads. The repartition of these clones significantly differed between South Australia and the rest of Australia. In addition, a significantly higher prevalence of 3/13 clones was observed in South Australia counties of the Mornington Peninsula, Melbourne and Bellarine Peninsula than in other South Australia counties. The data presented in this report suggest that the microevolution of three M. ulcerans complex clones drove the emergence of severe Buruli ulcer cases in some South Australia counties. Sequencing one specific PPE gene served to efficiently probe M. ulcerans complex clones. Further functional studies may balance the environmental adaptation and virulence of these clones.


2020 ◽  
Vol 117 (49) ◽  
pp. 31500-31509
Author(s):  
Shunsuke Watanabe ◽  
Naoki Takahashi ◽  
Yuri Kanno ◽  
Hiromi Suzuki ◽  
Yuki Aoi ◽  
...  

Active membrane transport of plant hormones and their related compounds is an essential process that determines the distribution of the compounds within plant tissues and, hence, regulates various physiological events. Here, we report that theArabidopsisNITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER FAMILY 7.3 (NPF7.3) protein functions as a transporter of indole-3-butyric acid (IBA), a precursor of the major endogenous auxin indole-3-acetic acid (IAA). When expressed in yeast, NPF7.3 mediated cellular IBA uptake. Loss-of-functionnpf7.3mutants showed defective root gravitropism with reduced IBA levels and auxin responses. Nevertheless, the phenotype was restored by exogenous application of IAA but not by IBA treatment.NPF7.3was expressed in pericycle cells and the root tip region including root cap cells of primary roots where the IBA-to-IAA conversion occurs. Our findings indicate that NPF7.3-mediated IBA uptake into specific cells is required for the generation of appropriate auxin gradients within root tissues.


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