Identification of a Novel Abscisic Acid-Regulated Farnesol Dehydrogenase from Arabidopsis

Bhandari, Jayaram; Fitzpatrick, A. Heather; Crowell, Dring N.

doi:10.1104/pp.110.157784

Cited by 23 publications

(27 citation statements)

References 38 publications

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“…We identified six of the seven enzymes of the mevalonate pathway as well as two enzymes involved in the sesquiterpenoid backbone biosynthesis (Fig. 5) [51,52]. In addition, we identified twelve proteins that are functionally related to the mevalonate pathway or have been reported as involved in pheromone or odor metabolism such as glutaminyl-tRNA synthetase, phosphoglycerate kinase, succinyl Co-A synthetase alpha subunit, transketolase, glutathione S transferases and cytochrome family proteins [53–56] (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Expression of the mevalonate pathway enzymes in the Lutzomyia longipalpis (Diptera: Psychodidae) sex pheromone gland demonstrated by an integrated proteomic approach

González-Caballero¹,

Rodríguez-Vega²,

Dias-Lopes³

et al. 2014

Journal of Proteomics

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In Latin America, Lutzomyia longipalpis is the main vector of the protozoan parasite Leishmania infantum, which is the causal agent of American Visceral Leishmaniasis. This insect uses male-produced pheromones for mate recognition. Elucidation of pheromone biogenesis or its regulation may enable molecular strategies for mating disruption and, consequently, the vector's population management. Motivated by our recent results of the transcriptomic characterization of the L. longipalpis pheromone gland, we performed a proteomic analysis of this tissue combining SDS-PAGE, and mass spectrometry followed by an integrative data analysis. Considering that annotated genome sequences of this sand fly are not available, we designed an alternative workflow searching MS/MS data against two customized databases using three search engines: Mascot, OMSSA and ProLuCID. A total of 542 proteins were confidently characterized, 445 of them using a Uniref100-insect protein database, and 97 using a transcript translated database. In addition, use of PEAKS for de novo peptide sequencing of MS/MS data confirmed ∼90% identifications made with the combination of the three search engines. Our results include the identification of six of the seven enzymes of the mevalonate-pathway, plus the enzymes involved in sesquiterpenoid biosynthesis, all of which are proposed to be involved in pheromone production in L. longipalpis. Biological significance L. longipalpis is the main vector of the protozoan parasite L. infantum, which is the causal agent of American Visceral Leishmaniasis. One of the control measures of such disease is focused on vector population control. As this insect uses male-produced pheromones for mate recognition, the elucidation of pheromone biogenesis or its regulating process may enable molecular strategies for mating disruption and, consequently, this vector's population management. On this regard, in this manuscript we report expression evidence, at the protein level, of several molecules potentially involved in the pheromone production of L. longipalpis. Our results include the identification of the mevalonate-pathway enzymes, plus the enzymes involved in sesquiterpenoid biosynthesis, all of which are proposed to be involved in pheromone production in L. longipalpis. In addition, considering that the annotated genome sequences of this sand fly are not yet available, we designed an alternative workflow searching MS/MS data against proteomic and transcript translated customized databases, using three search engines: Mascot, OMSSA, and ProLuCID. In addition, a de novo peptide sequencing software (PEAKS) was used to further analyze the MS/MS data. This approach made it possible to identify and annotate 542 proteins for the pheromone gland of L. longipalpis. Importantly, all annotated protein sequences and raw data are available for the research community in protein repositories that provide free access to the data.

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Section: Resultsmentioning

confidence: 99%

Expression of the mevalonate pathway enzymes in the Lutzomyia longipalpis (Diptera: Psychodidae) sex pheromone gland demonstrated by an integrated proteomic approach

González-Caballero¹,

Rodríguez-Vega²,

Dias-Lopes³

et al. 2014

Journal of Proteomics

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“…A number of genes involved in isoprenoid metabolism and ABA signaling are ABA-regulated (Huizinga et al, 2008;Bhandari et al, 2010). To determine whether the FOLK gene is similarly regulated by ABA, the effect of exogenous ABA on FOLK expression was analyzed.…”

Section: Aba Regulation Of Folk Gene Expressionmentioning

confidence: 99%

“…RT-PCR fragments were resolved by agarose gel electrophoresis, visualized by ethidium bromide staining, inserted into pYES2.1/V5-His-TOPO according to the manufacturer's instructions (Invitrogen/Life Technologies), and analyzed on an Applied Biosystems 3130XL genetic analyzer (http:// www.appliedbiosystems.com/) to confirm DNA sequences and insert orientations. Recombinant plasmids were then introduced into Saccharomyces cerevisiae strain SM1058 (MATa leu2, ura3, trp1, his4, can1) as described previously (Bhandari et al, 2010). Transformed yeast cells were selected on CSM-ura agar medium, colony-purified, and grown at 30°C in CSM-ura liquid medium containing 2% glucose.…”

Section: Farnesol Kinase Assaysmentioning

confidence: 99%

“…Because other genes involved in farnesol metabolism have been shown to influence ABA signaling (Bhandari et al, 2010), folk mutants were analyzed for altered ABA signaling. All three folk mutants were found to exhibit a hypersensitive response to ABA in seed germination assays, establishing the product of the FOLK gene as a negative regulator of ABA signaling.…”

Section: Aba Regulation Of Folk Gene Expressionmentioning

confidence: 99%

“…Because other genes involved in farnesol metabolism have an ABA phenotype (Bhandari et al, 2010), the folk mutants characterized in Figure 6 were analyzed for ABA responsiveness in seed germination assays. As shown in Figure 7, folk-1, -2 and -3 exhibited increased inhibition of seed germination in the presence of exogenous ABA.…”

Section: Phenotypic Analysis Of Folk Loss-of-function Mutantsmentioning

confidence: 99%

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Farnesol kinase is involved in farnesol metabolism, ABA signaling and flower development in Arabidopsis

2011

Self Cite

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SUMMARYFarnesol, which is toxic to plant cells at high concentrations, is sequentially phosphorylated to farnesyl phosphate and farnesyl diphosphate. However, the genes responsible for the sequential phosphorylation of farnesol have not been identified and the physiological role of farnesol phosphorylation has not been fully elucidated. To address these questions, we confirmed the presence of farnesol kinase activity in Arabidopsis (Arabidopsis thaliana) membranes and identified the corresponding gene (At5g58560, FOLK). Heterologous expression in recombinant yeast cells established farnesol as the preferred substrate of the FOLK-encoded kinase. Moreover, loss-of-function mutations in the FOLK gene abolished farnesol kinase activity, caused an abscisic acid-hypersensitive phenotype and promoted the development of supernumerary carpels under water-stress conditions. In wild-type plants, exogenous abscisic acid repressed FOLK gene expression. These observations demonstrate a role for farnesol kinase in negative regulation of abscisic acid signaling, and provide molecular evidence for a link between farnesol metabolism, abiotic stress signaling and flower development.

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Biosynthesis of geranate via isopentenol utilization pathway in Escherichia coli

Pan

Hong

et al. 2022

Biotech & Bioengineering

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Isoprenoids are a large family of natural products with diverse structures, which allow them to play diverse and important roles in the physiology of plants and animals. They also have important commercial uses as pharmaceuticals, flavoring agents, fragrances, and nutritional supplements. Recently, metabolic engineering has been intensively investigated and emerged as the technology of choice for the production of isoprenoids through microbial fermentation. Isoprenoid biosynthesis typically originates in plants from acetyl‐coA in central carbon metabolism, however, a recent study reported an alternative pathway, the isopentenol utilization pathway (IUP), that can provide the building blocks of isoprenoid biosynthesis from affordable C5 substrates. In this study, we expressed the IUP in Escherichia coli to efficiently convert isopentenols into geranate, a valuable isoprenoid compound. We first established a geraniol‐producing strain in E. coli that uses the IUP. Then, we extended the geraniol synthesis pathway to produce geranate through two oxidation reactions catalyzed by two alcohol/aldehyde dehydrogenases from Castellaniella defragrans. The geranate titer was further increased by optimizing the expression of the two dehydrogenases and also parameters of the fermentation process. The best strain produced 764 mg/L geranate in 24 h from 2 g/L isopentenols (a mixture of isoprenol and prenol). We also investigated if the dehydrogenases could accept other isoprenoid alcohols as substrates.

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Identification of a Novel Abscisic Acid-Regulated Farnesol Dehydrogenase from Arabidopsis

Cited by 23 publications

References 38 publications

Expression of the mevalonate pathway enzymes in the Lutzomyia longipalpis (Diptera: Psychodidae) sex pheromone gland demonstrated by an integrated proteomic approach

Expression of the mevalonate pathway enzymes in the Lutzomyia longipalpis (Diptera: Psychodidae) sex pheromone gland demonstrated by an integrated proteomic approach

Farnesol kinase is involved in farnesol metabolism, ABA signaling and flower development in Arabidopsis

Biosynthesis of geranate via isopentenol utilization pathway in Escherichia coli

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