The VEP1 gene (At4g24220) encodes a short-chain dehydrogenase/reductase with 3-oxo-Δ4,5-steroid 5β-reductase activity in Arabidopsis thaliana L.

Herl, Vanessa; Fischer, Gabriele; Reva, Veaceslav; Stiebritz, Martin T.; Muller, Yves A.; Müller‐Uri, Frieder; Kreis, Wolfgang

doi:10.1016/j.biochi.2008.12.005

Cited by 39 publications

(38 citation statements)

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“…euphratica was confirmed to play key roles in salt and drought stress tolerance [51]. In the present study, Ptc-miR473 was confirmed to be targeted to Vein Patterning 1 (VEP1), which belongs to a short-chain dehydrogenase/reductase (SDR) superfamily [52]. The homolog of VEP1 in Arabidopsis was confirmed to be required for vascular strand development and to be upregulated by osmotic stress [52,53].…”

Section: Discussionmentioning

confidence: 76%

“…In the present study, Ptc-miR473 was confirmed to be targeted to Vein Patterning 1 (VEP1), which belongs to a short-chain dehydrogenase/reductase (SDR) superfamily [52]. The homolog of VEP1 in Arabidopsis was confirmed to be required for vascular strand development and to be upregulated by osmotic stress [52,53]. Ptc-miR473 regulates the expression the GRAS protein and VEP1, both of which were responsive to drought stress, this may be the drought tolerance mechanism in P .…”

Section: Discussionmentioning

confidence: 99%

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Identification of drought-responsive and novel Populus trichocarpamicroRNAs by high-throughput sequencing and their targets using degradome analysis

et al. 2013

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BackgroundMicroRNAs (miRNAs) are endogenous small RNAs (sRNAs) with a wide range of regulatory functions in plant development and stress responses. Although miRNAs associated with plant drought stress tolerance have been studied, the use of high-throughput sequencing can provide a much deeper understanding of miRNAs. Drought is a common stress that limits the growth of plants. To obtain more insight into the role of miRNAs in drought stress, Illumina sequencing of Populus trichocarpa sRNAs was implemented.ResultsTwo sRNA libraries were constructed by sequencing data of control and drought stress treatments of poplar leaves. In total, 207 P. trichocarpa conserved miRNAs were detected from the two sRNA libraries. In addition, 274 potential candidate miRNAs were found; among them, 65 candidates with star sequences were chosen as novel miRNAs. The expression of nine conserved miRNA and three novel miRNAs showed notable changes in response to drought stress. This was also confirmed by quantitative real time polymerase chain reaction experiments. To confirm the targets of miRNAs experimentally, two degradome libraries from the two treatments were constructed. According to degradome sequencing results, 53 and 19 genes were identified as targets of conserved and new miRNAs, respectively. Functional analysis of these miRNA targets indicated that they are involved in important activities such as the regulation of transcription factors, the stress response, and lipid metabolism.ConclusionsWe discovered five upregulated miRNAs and seven downregulated miRNAs in response to drought stress. A total of 72 related target genes were detected by degradome sequencing. These findings reveal important information about the regulation mechanism of miRNAs in P. trichocarpa and promote the understanding of miRNA functions during the drought response.

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Section: Discussionmentioning

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Identification of drought-responsive and novel Populus trichocarpamicroRNAs by high-throughput sequencing and their targets using degradome analysis

et al. 2013

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“…Solvents were purchased from Carl Roth GmbH (Karlsruhe, Germany). Successful enzymatic conversion of progesterone (0.3 mM) to 5ß-pregnane-3,20-dione was achieved at 40°C in samples containing 0.2 mg/mL recombinant protein and a NADPH regenerating system, consisting of 6.4 mM NADP, 32.1 mM glucose-6-phosphate, and 42 nkat glucose-6-phosphate dehydrogenase as described by Herl et al (2009). Heat-inactivated samples (10 min, 100°C) served as controls.…”

Section: Enzymatic Tests Including Product Identificationmentioning

confidence: 99%

“…Related enzymes have been detected in other plant species that do not accumulate cardenolides, following the initial purification of P5ßR activity from Digitalis purpurea L. by Gärtner et al 1994. However, the identification of the resulting biochemical products remains a challenge (Herl et al 2006(Herl et al , 2009Gavidia et al 2007;Bauer et al 2010;Perez-Bermudez et al 2010;Munkert et al 2011Munkert et al , 2015aMunkert et al , 2015Ernst et al 2015).…”

Section: Introductionmentioning

confidence: 99%

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The occurrence of progesterone 5β-reductase is not limited to the angiosperms: a functional gene was identified in Picea sitchensis and expressed in Escherichia coli

Rudolph

Wiegert

Schubert

et al. 2016

N.Z. j. of For. Sci.

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Background: Progesterone 5β-reductases (P5ßRs) catalyse one step in the stereospecific biosynthesis of cardenolides (potent repellents of herbivores and pharmaceutical stimulants of disordered heart muscle cells). They were originally discovered in the genus Digitalis and have since been frequently isolated from other angiosperms. Recombinant P5ßRs engineered in Escherichia coli host cells convert a broad spectrum of compounds in vitro, sometimes with higher efficiency than with progesterone. This observation suggests additional functions for cardenolide metabolism and promises future use in sustainable chemistry and biotechnology. Methods: A tissue complementary DNA (cDNA) library was screened for orthologous P5ßRs. Candidates were subcloned into expression vectors and overexpressed in E. coli cells. The recombinant P5ßR protein was investigated for catalytic activity with several related substrates. Using spectrophotometric assays, the biochemical parameters of the enzyme were calculated. A 3D model was created and was compared to the previously published P5ßR structure of Digitalis lanata and other plant P5ßR models.Results: Performing protein similarity searches in public databases and comparison of 3D protein structure models revealed four cDNA clones in a tissue library of Picea sitchensis (Bong.) Carrière putatively encoding P5ßRs. Succeeding with the expression of one clone in E. coli, the highly purified protein was unambiguously able to enantioselectively convert progesterone into 5ß-pregnane-3,20-dione. However, the catalytic activity to reduce the small molecule 2-cyclohexen-1-one was nearly 100 times faster. Methyl vinyl ketone was reduced similar to results from previously studied angiosperm resources.Conclusions: (i) The low catalytic efficiency for progesterone conversion agrees with the fact that conifers have not been reported to accumulate cardenolides. This finding suggests that alternate metabolic processes occur whereby the newly detected enzymes could transform smaller molecules rather than large ones such as progesterone.(ii) An ancient P5ßR gene appears to have existed in the last common ancestor of seed plants approximately 300 million years ago. If the diversification of P5ßRs, including the currently detected homologous iridoid synthase activity, was related to stress encountered during the transition to growth on land, then investigation of P5ßRs from pteridophytes and bryophytes should improve our knowledge of this enzyme class and elucidate the direction of evolution.

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Synthetic Strategies Based on CC Bioreductions for the Preparation of Biologically Active Molecules

Gatti¹,

Parmeggiani²,

Sacchetti³

2013

Synthetic Methods for Biologically Active Molecules

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The VEP1 gene (At4g24220) encodes a short-chain dehydrogenase/reductase with 3-oxo-Δ4,5-steroid 5β-reductase activity in Arabidopsis thaliana L.

Cited by 39 publications

References 36 publications

Identification of drought-responsive and novel Populus trichocarpamicroRNAs by high-throughput sequencing and their targets using degradome analysis

Identification of drought-responsive and novel Populus trichocarpamicroRNAs by high-throughput sequencing and their targets using degradome analysis

The occurrence of progesterone 5β-reductase is not limited to the angiosperms: a functional gene was identified in Picea sitchensis and expressed in Escherichia coli

Synthetic Strategies Based on CC Bioreductions for the Preparation of Biologically Active Molecules

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