Molecular cloning and characterization of the <i>Arabidopsis thaliana</i> α‐subunit of elongation factor 1B

Héricourt, François; Jupin, Isabelle

doi:10.1016/s0014-5793(99)01694-4

Cited by 6 publications

(6 citation statements)

References 27 publications

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“…An elongation factor 1B alpha-subunit 2 protein (eEF1Balpha2; spot 406) was up-regulated in Arabidopsis after salt treatment (Table ). It catalyzes the GTP-dependent binding of aminoacyl-tRNA to the aminoacyl site on ribosomes concomitant with the hydrolysis of GTP . It is postulated that enhancement of eEF1Balpha2 protein in salt stress serves to maintain the normal operation of protein synthesis.…”

Section: Resultssupporting

confidence: 80%

Comparative Proteomics of Salt Tolerance in Arabidopsis thaliana and Thellungiella halophila

et al. 2010

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Salinity is a major abiotic stress affecting plant cultivation and productivity. Thellungiella halophila is a halophyte and has been used as a model for studying plant salt tolerance. Understanding the molecular mechanisms of salinity tolerance will facilitate the generation of salt tolerant crops. Here we report comparative leaf proteomics of Arabidopsis, a glycophyte, and its close relative Thellungiella, a halophyte, under different salt stress conditions. Proteins from control and NaCl treated Arabidopsis and Thellungiella leaf samples were extracted and separated by two-dimensional gel electrophoresis. A total of 88 protein spots from Arabidopsis gels and 37 protein spots from Thellungiella gels showed significant changes. Out of these spots, a total of 79 and 32 proteins were identified by mass spectrometry in Arabidopsis and Thellungiella, respectively. Most of the identified proteins were involved in photosynthesis, energy metabolism, and stress response in Arabidopsis and Thellungiella. As a complementary approach, isobaric tag for relative and absolute quantification (iTRAQ) LC-MS was used to identify crude microsomal proteins. A total of 31 and 32 differentially expressed proteins were identified in Arabidopsis and Thellungiella under salt treatment, respectively. Overall, there were more proteins changed in abundance in Arabidopsis than in Thellungiella. Distinct patterns of protein changes in the two species were observed. Collectively, this work represents the most extensive proteomic description of salinity responses of Arabidopsis and Thellungiella and has improved our knowledge of salt tolerance in glycophytes and halophytes.

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Section: Resultssupporting

confidence: 80%

Comparative Proteomics of Salt Tolerance in Arabidopsis thaliana and Thellungiella halophila

et al. 2010

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“…Both cDNAs were found to contain full-length coding regions. The putative product of the CPHO62 cDNA (989 bp) was found to be a protein elongation factor of the 1B class (66% identity with the corresponding proteins from Arabidopsis thaliana and rice; Hericourt and Jupin 1999). More specifically, the open reading frame (ORF) detected between positions 70 and 612 (corresponding to a polypeptide of 180 amino acids) is predicted to encode the GDP-GTP nucleotide-exchange subunit of the translation elongation complex.…”

Section: Methylation-dependent Markers Of Embryogenic Callus Linesmentioning

confidence: 99%

Methylation-sensitive RFLPs: characterisation of two oil palm markers showing somaclonal variation-associated polymorphism

2002

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The occurrence of "mantled" somaclonal variants (approx. 5%), which display alterations in floral organ structure, among populations of somatic embryo-derived oil palms ( Elaeis guineensis Jacq.) currently hampers any scaling-up of clonal plant micropropagation. As a first step towards the identification of abnormality-discriminating markers, we have screened a set of 27 oil palm cDNA probes for methylation-sensitive restriction fragment length polymorphisms (RFLPs) using callus genomic DNA digested with the isoschizomeric enzymes MspI and HpaII. Only two probes (CPHO62 and -63) were found to differentiate reproducibly in two different genotypic backgrounds between nodular compact calli (NCC) and fast-growing calli (FGC), which generate 5% and 100% "mantled" plantlets, respectively. Comparative analyses were then conducted on DNA from inflorescences and leaves of normal and abnormal adult regenerants. With both probes, the observed methylation patterns were strongly clone-dependent and monomorphic with respect to the phenotype of the regenerants, except for the type-specific banding pattern obtained with the CPHO62 probe on material from the LMC3 clonal offspring. The results presented here mirror the higher difference in genomic DNA methylation observed between normal and abnormal embryogenic calli when compared to more differentiated plant material. Moreover, they reinforce the paramount interest of NCC and FGC callus lines as a material of choice in the search for early epigenetic markers of the "mantled" somaclonal variation. The potential use of methylation-sensitive RFLPs for the early detection of somaclonal variation at early stages of the micropropagation process is discussed.

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“…Plasmids pNS10, pNtYI, and pGNPS include tobacco (Nicotiana tabacum) promoters from rbcS (Ntss23), tRNA Tyr , and b-1,3-glucanase genes, respectively (Mazur and Chui, 1985;Stange and Beier, 1986;Yukawa et al, 2001). Plasmids pYY0847 and pYI0805 contain just the CaMV 35S promoter and the CaMV 35S promoter fused to part of the coding region of Arabidopsis elongation factor 1Ba1 (Héricourt and Jupin, 1999), respectively. Plasmid pRBC-1657 includes the promoter from the tomato (Solanum lycopersicum) rbcS gene for details, see Supplemental Fig.…”

Section: Plasmids For In Vitro Transcription Assaysmentioning

confidence: 99%

Arabidopsis Pol II-Dependent in Vitro Transcription System Reveals Role of Chromatin for Light-InduciblerbcSGene Transcription

Ido

Iwata

et al. 2015

Plant Physiology

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In vitro transcription is an essential tool to study the molecular mechanisms of transcription. For over a decade, we have developed an in vitro transcription system from tobacco (Nicotiana tabacum)-cultured cells (BY-2), and this system supported the basic activities of the three RNA polymerases (Pol I, Pol II, and Pol III). However, it was not suitable to study photosynthetic genes, because BY-2 cells have lost their photosynthetic activity. Therefore, Arabidopsis (Arabidopsis thaliana) in vitro transcription systems were developed from green and etiolated suspension cells. Sufficient in vitro Pol II activity was detected after the minor modification of the nuclear soluble extracts preparation method; removal of vacuoles from protoplasts and L-ascorbic acid supplementation in the extraction buffer were particularly effective. Surprisingly, all four Arabidopsis Rubisco small subunit (rbcS-1A, rbcS-1B, rbcS-2B, and rbcS-3B) gene members were in vitro transcribed from the naked DNA templates without any light-dependent manner. However, clear light-inducible transcriptions were observed using chromatin template of rbcS-1A gene, which was prepared with a human nucleosome assembly protein 1 (hNAP1) and HeLa histones. This suggested that a key determinant of light-dependency through the rbcS gene transcription was a higher order of DNA structure (i.e. chromatin).In plants, the molecular mechanisms of lightdependent gene regulation remain largely unknown. Generally, transcription is the major regulation step during gene expression in the eukaryotic genome. In plants, most genes are mainly regulated at the level of transcription, and the expressions of many genes depend on surrounding light conditions relating to photosynthesis, photomorphogenesis, phototropism, circadian rhythms, gametogenesis, etc. Therefore, light-dependent transcription has been studied intensively. Three distinct classes of photoreceptors have been discovered: phytochromes, blue-light receptors (cryptochromes, phototropins, and ZTL / FKF1 / LKP2 family), and photoreceptors for UV-B (Banerjee and Batschauer, 2005). These photoreceptors affect downstream gene expression mediated by the specific binding of transcription factors to cis-regulatory elements in promoters. Light regulatory elements are essential cisregulatory elements found in photo-responsible genes, such as the conserved modular array 5 (CMA5) on the Rubisco small subunit (rbcS) genes (López-Ochoa et al., 2007). Our knowledge of transacting factors that associate with light regulatory elements is quite limited, mainly because plant materials are inconvenient to use for biochemical research. Therefore, to understand transcription regulation comprehensively, we need to develop a plant-specific in vitro transcription system.The eukaryotic (human cell) in vitro transcription system was reported by Weil et al. (1979) and Manley et al. (1980). Subsequently, Dignam et al. (1983) established finely tuned systems. Nowadays, in vitro transcription systems are indispensable for molecular biology;...

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Molecular cloning and characterization of the Arabidopsis thaliana α‐subunit of elongation factor 1B

Cited by 6 publications

References 27 publications

Comparative Proteomics of Salt Tolerance in Arabidopsis thaliana and Thellungiella halophila

Comparative Proteomics of Salt Tolerance in Arabidopsis thaliana and Thellungiella halophila

Methylation-sensitive RFLPs: characterisation of two oil palm markers showing somaclonal variation-associated polymorphism

Arabidopsis Pol II-Dependent in Vitro Transcription System Reveals Role of Chromatin for Light-InduciblerbcSGene Transcription

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