Light Induction of Translatable mRNAs for Chloroplastic Leucyl- and Valyl-tRNA Synthetases of Euglena gracilis

Krauspe, R.; Lerbs, S.; Parthier, B.; Wollgiehn, R.

doi:10.1016/s0176-1617(87)80199-2

Cited by 19 publications

(12 citation statements)

References 55 publications

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“…mmol Amersham International PLC, England), RNA amounts were used that gave at least 65% of maximal incorporation of radioactivity Cultivation of E. gracilis into trichloroacetic-acid-precipitable material. The transcollection, Gottingen) were cultivated photoautotrophically lation products were analysed by two-dimensional gel as described [19]. electrophoresis as described above.…”

Section: Methodsmentioning

confidence: 99%

“…For the determination of 5-enolpyruvylshikimate-3-phosphate synthase activity after non-denaturing PAGE [19,221, two consecutive 1-mm gel slices from lanes separating 1 pg protein from adapted and control cells were incubated in 100-p1 assays containing 10 mM shikimate 3-phosphate, 3.25 mM potassium fluoride, 65 mM Hepes and 1 mM phosphoenolpyruvate at 37°C for 5 min. After boiling to terminate the reaction, the remaining phosphoenolpyruvate was determined as above.…”

Section: Electrophoretic Techniquesmentioning

confidence: 99%

“…Preparation of RNA After phenol/chloroform extraction from deep-frozen cells, high-molecular-mass RNA was reprecipitated with 3 M LiCl in 97.5 mM NaCl and 9.75 mM sodium citrate, pH 7.0, as described in [19].…”

Section: Other Determinationsmentioning

confidence: 99%

“…In vitro translation of RNA Variable amounts of RNA dissolved in tridistilled water were used to direct protein synthesis in a cell-free wheat Wild-type photoautotrophic Euglena cells responded to Ngerm system [19]. (Fig.…”

Section: Other Determinationsmentioning

confidence: 99%

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N‐(Phosphonomethyl)glycine (glyphosate) tolerance in Euglena gracilis acquired by either overproduced or resistant 5‐enolpyruvylshikimate‐3‐phosphate synthase

Reinbothe

Nelles

Parthier

1991

European Journal of Biochemistry

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Photoautotrophic cells of Euglena gracilis can be adapted to N-(phosphonomethy1)glycine (glyphosate) by cultivation in media with progressively higher concentrations of the herbicide. Two different mechanisms of tolerance to the herbicide were observed. One is characterized by the overproduction and 40-fold accumulation of the target enzyme, 5-enolpyruvylshikimate-3-phosphate synthase, in cells adapted to 6 mM N-(phosphonomethy1)glycine. The other is connected with a herbicide-insensitive enzyme. No evidence was obtained for the involvement of the putative multifunctional arom protein previously reported to be involved in the biosynthesis of aromatic amino acids in Euglena. Cells adapted to N-(phosphonomethy1)glycine excreted shikimate and shikimate 3-phosphate into the medium; the amounts depended on the actual concentration of the herbicide. Two-dimensional gel electrophoresis and determination of 5-enolpyruvylshikimate-3-phosphate synthase activity in crude extracts, as well as after separation by non-denaturing gel electrophoresis, revealed that the overproduction of the enzyme in adapted cells correlates with the accumulation of a 59-kDa protein. Overproduction of this 59-kDa protein resulted from a selectively increased level of a mRNA coding for a 64.5-kDa polypeptide which appeared in adapted cells, as shown by cell-free translation in the wheat germ system.In contrast to this quantitative, adaptive type of tolerance, the second mechanism causing tolerance to N-(phosphonomethy1)glycine in the Euglena cell line NR 6/50 was probably related to a qualitatively altered 5-enolpyruvylshikimate-3-phosphate synthase, which could not be inhibited by even 2 mM N-(phosphonomethy1)glycine in vitro. In agreement with this observation, the putatively mutated cell line excreted neither shikimate nor shikimate 3-phosphate into the growth medium containing N-(phosphonomethyl)glycine, even if cultivated in the presence of 20 mM or 50 mM N-(phosphonomethy1)glycine. synthases characterized so far are monomeric plastid proteins of a relatively uniform molecular mass between 46-kDa and 47.6-kDa.In contrast to higher plants and bacteria, S-enolpyruvylshikimate-3-phosphate synthases of higher, as well as several lower fungi, e. g. from Neurospora crassa [l 1, 121, Aspergillus nidulans [13] and Schizosaccharomycespombe [14,15], are part of the multifunctional arom protein. A similar organization of enzyme activities, which convert 3-deoxy-~-arabinoheptulosonate 7-phosphate to 5-enolpyruvylshikimate 3-phosphate by steps 2 -6 of the pre-chorismate pathway, has been proposed for the unicellular phytoflagellate Euglena gracilis [16, 171. Since it is known that the 5-enolpyruvylshikimate-3-phosphate synthase domain of the pentafunctional arom protein of N. crassa is sensitive to N-(phosphonomethy1)glycine [18], this should be an excellent tool for probing the nature of the arom protein in E. gracilis.In this paper we demonstrate that Euglena can adapt to N-(phosphonomethy1)glycine when the herbicide concentration of the medium is p...

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Section: Methodsmentioning

confidence: 99%

Section: Electrophoretic Techniquesmentioning

confidence: 99%

Section: Other Determinationsmentioning

confidence: 99%

Section: Other Determinationsmentioning

confidence: 99%

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N‐(Phosphonomethyl)glycine (glyphosate) tolerance in Euglena gracilis acquired by either overproduced or resistant 5‐enolpyruvylshikimate‐3‐phosphate synthase

Reinbothe

Nelles

Parthier

1991

European Journal of Biochemistry

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“…Light induced increases in the specific activity of the nuclear encoded chloroplast initiation factors IF1, IF2, IF3, (Gold and Spremulli 1985;Spremulli 1986, 1988), the nuclear encoded chloroplast elongation factors EF-Ts, EF-G (Breitenberger et al 1979;Eberly et al 1986;Fox et al 1980) and the chloroplast encoded chloroplast elongation factor EF-Tu (Eberly et al 1986;Montandon and Stutz 1983;Passavant et al 1983;Spremulli 1982) have also been reported. The synthesis of the nuclear encoded chloroplast localized aminoacyltRNA synthetases (Hecker et al 1974;Krauspe et al 1987;McCarthy et al 1982;Nover 1976;Parthier and Krauspe 1974;Parthier et al 1972;Reger et al 1970;Schimpf et al 1982) is induced by light exposure. The synthesis of chloroplast rRNA (Cohen and Schiff 1976) and chloroplast aminoacyl-tRNA synthetases (McCarthy et al 1982) continues when cells are returned to the dark indicating that induction is a light triggered process.…”

Section: Photocontrol Of Chloroplast Developmentmentioning

confidence: 99%

Photo and Nutritional Regulation of Euglena Organelle Development

Schwartzbach

2017

Advances in Experimental Medicine and Biology

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Euglena can use light and CO, photosynthesis, as well as a large variety of organic molecules as the sole source of carbon and energy for growth. Light induces the enzymes, in this case an entire organelle, the chloroplast, that is required to use CO as the sole source of carbon and energy for growth. Ethanol, but not malate, inhibits the photoinduction of chloroplast enzymes and induces the synthesis of the glyoxylate cycle enzymes that comprise the unique metabolic pathway leading to two carbon, ethanol and acetate, assimilation. In resting, carbon starved cells, light mobilizes the degradation of the storage carbohydrate paramylum and transiently induces the mitochondrial proteins required for the aerobic metabolism of paramylum to provide the carbon and energy required for chloroplast development. Other mitochondrial proteins are degraded upon light exposure providing the amino acids required for the synthesis of light induced proteins. Changes in protein levels are due to increased and decreased rates of synthesis rather than changes in degradation rates. Changes in protein synthesis rates occur in the absence of a concomitant increase in the levels of mRNAs encoding these proteins indicative of photo and metabolic control at the translational rather than the transcriptional level. The fraction of mRNA encoding a light induced protein such as the light harvesting chlorophyll a/b binding protein of photosystem II, (LHCPII) associated with polysomes in the dark is similar to the fraction associated with polysomes in the light indicative of photoregulation at the level of translational elongation. Ethanol, a carbon source whose assimilation requires carbon source specific enzymes, the glyoxylate cycle enzymes, represses the synthesis of chloroplast enzymes uniquely required to use light and CO as the sole source of carbon and energy for growth. The catabolite sensitivity of chloroplast development provides a mechanism to prioritize carbon source utilization. Euglena uses all of its resources to develop the metabolic capacity to utilize carbon sources such as ethanol which are rarely in the environment and delays until the rare carbon source is no longer available forming the chloroplast which is required to utilize the ubiquitous carbon source, light and CO.

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Cellular Differentiation Processes During Chloroplast Development in Euglena Gracilis

Reinbothe

1992

Regulation of Chloroplast Biogenesis

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Light Induction of Translatable mRNAs for Chloroplastic Leucyl- and Valyl-tRNA Synthetases of Euglena gracilis

Cited by 19 publications

References 55 publications

N‐(Phosphonomethyl)glycine (glyphosate) tolerance in Euglena gracilis acquired by either overproduced or resistant 5‐enolpyruvylshikimate‐3‐phosphate synthase

N‐(Phosphonomethyl)glycine (glyphosate) tolerance in Euglena gracilis acquired by either overproduced or resistant 5‐enolpyruvylshikimate‐3‐phosphate synthase

Photo and Nutritional Regulation of Euglena Organelle Development

Cellular Differentiation Processes During Chloroplast Development in Euglena Gracilis

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