Analysis of the cucumber malate synthase gene promoter by transient expression and gel retardation assays

Graham, Ian A.; Baker, Christopher J. O.; Leaver, Christopher J.

doi:10.1046/j.1365-313x.1994.6060893.x

Cited by 33 publications

(28 citation statements)

References 27 publications

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“…Promoter analyses of sugar-starvation-inducible genes, such as malate synthase (Graham et al 1994, Sarah et al 1996, a-amylase (Hwang et al 1998, Lu et al 1998, Toyofuku et al 1998) and a photosynthetic gene (Urwin and Jenkins 1997) have identified several cis-regulatory sequences. However, none of these putative cis-regulatory elements was found in the promoter regions of Arabidopsis E1b and E2 genes.…”

Section: Discussionmentioning

confidence: 99%

Activation of the Promoters of Arabidopsis Genes for the Branched-Chain α-Keto Acid Dehydrogenase Complex in Transgenic Tobacco BY-2 Cells under Sugar Starvation

Fujiki¹,

Ito²,

Itoh³

et al. 2002

Plant and Cell Physiology

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;Sugar starvation exerted by sub-10 mM levels of sucrose on Arabidopsis T87 suspension-cultured cells triggered marked accumulation of the transcripts of genes for E1> > > > and E2 subunit of the branched-chain = = = =-keto acid dehydrogenase complex. Similar levels of sugar starvation increased the luciferase activity in transgenic tobacco BY-2 lines expressing the Arabidopsis E1> > > >-or E2-promoterluciferase fusion gene. These results indicate that sugar levels tightly regulate the E1> > > > and E2 promoter activity in the heterologous plant system. We further showed in the transgenic tobacco BY-2 lines that sugar-starvation-induced activation of the E1> > > > and E2 promoters was prevented by K-252a, an inhibitor of Ser/Thr protein kinase, and was enhanced by okadaic acid, an inhibitor of protein phosphatases. By contrast, the cauliflower mosaic virus 35S promoter activity in sugar-starved BY-2 cells was not significantly affected by K-252a and only slightly enhanced by okadaic acid. Taken together, we propose that transcriptional activation of genes for the branched-chain = = = =-keto acid dehydrogenase complex and its modulation by specific protein kinases/phosphatases are of critical importance in branched-chain amino acid catabolism in plant cells under sugar starvation.

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Section: Discussionmentioning

confidence: 99%

Activation of the Promoters of Arabidopsis Genes for the Branched-Chain α-Keto Acid Dehydrogenase Complex in Transgenic Tobacco BY-2 Cells under Sugar Starvation

Fujiki¹,

Ito²,

Itoh³

et al. 2002

Plant and Cell Physiology

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“…In our cucumber study, two highly conserved germination-response elements (IMHI and IMH2) were identified in the CsPCK gene as being necessary for germination and, possibly, senescence. Although the biological function of the IMHI sequence is not yet clear, similar sequences have been shown in fungi (Neurospora crassa) and yeast (Saccharomyces cerevisiae), suggesting that it may be required for either a sugar response (starvation) or germination response (Graham et al, 1994). The IMH2 sequences (with similarity to MYB binding sequences) in the icl, ms, and CsPCK promoters might direct the carbohydrate regulation of expression for many plant genes (Graham eta[., 1994;Grotewold et al, 1994).…”

Section: Discussionmentioning

confidence: 87%

“…Therefore, controlling intracellular sugar levels is a central element for the regulation of metabolic pathways in living cells. Many of the genes associated with carbon assimilation and sugar transport are directly or partly regulated by fluxes in the sugar levels of target cells and tissues (Sheen, 1990;Graham et al, 1994;Koch, 1996;Sherson et al, 2000). Because of this, the metabolic regulation of plant gene expression is now a central focus in studying the plant sugartransport mechanism.…”

Section: Discussionmentioning

confidence: 99%

Molecular cloning and structural analysis of the cucumber (Cucumis sativus L.) phosphoenolpyruvate carboxykinase (CsPCK) gene

Kim

2003

J. Plant Biol.

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“…Others have shown that high levels of metabolizable carbohydrates can cause repression in the transcription of these genes in cultured plant cells (Kudielka et al, 1981;Theimer, 1983a, 1983b;Graham et al, 1992Graham et al, , 1994a. Because distinct DNA sequences play roles in activating the isocitrate lyase gene in developing seed and seedlings, it seems unlikely that its expression is controlled solely by carbohydrate levels.…”

Section: Several Dna Sequences Regulate the Lsocitrate Lyase Gene Durmentioning

confidence: 99%

DNA Sequences That Activate Isocitrate Lyase Gene Expression during Late Embryogenesis and during Postgerminative Growth

et al. 1996

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We analyzed DNA sequences that regulate the expression of an isocitrate lyase gene from Brasska napus L. during late embryogenesis and during postgerminative growth to determine whether glyoxysomal function is induced by a common mechanism at different developmental stages. P-Clucuronidase constructs were used both in transient expression assays in 6. napus and in transgenic Arabidopsis thaliana to identify the segments of the isocitrate lyase 5' flanking region that influence promoter activity. DNA sequences that play the principal role in activating the promoter during postgerminative growth are located more than 1200 bp upstream of the gene. Distinct DNA sequences that were sufficient for high-leve1 expression during late embryogenesis but only low-leve1 expression during postgerminative growth were also identified. Other parts of the 5' flanking region increased promoter activity both in developing seed and in seedlings. We conclude that a combination of elements is involved in regulating the isocitrate lyase gene and that distinct DNA sequences play primary roles in activating the gene in embryos and in seedlings. These findings suggest that different signals contribute to the induction of glyoxysomal function during these two developmental stages. We also showed that some of the constructs were expressed differently in transient expression assays and in transgenic plants.Glyoxysomes are peroxisomes with a specialized function in lipid mobilization. They contain enzymes of the P-oxidation pathway and the glyoxylate cycle that largely account for the ability of plants to utilize lipids as a carbon source (Trelease and Doman, 1984). These organelles play a prominent role in the metabolism of postgerminative seedlings by converting storage lipids into carbohydrates that serve as carbon and energy sources for the growing seedling (Bridenbach et al., 1967;Huang et al., 1983;Trelease, 1984). Functional glyoxysomes are also present in senescing organs, where they are thought to play a similar role in transforming membrane lipids into carbohydrates that can This work was supported by grants from the National Science Foundation. C.M.S. was supported in part by a fellowship from the North Atlantic Treaty Organization. Bellis and Nishimura, 1991;Wanner et al., 1991;Graham et al., 1992). The findings that genes encoding the two glyoxysome-specific enzymes, isocitrate lyase and malate synthase, are active during pollen and seed development as well as during postgerminative growth opens the possibility that peroxisomes present at these stages of the life cycle may have a glyoxysomal function (Pais and Feijo, 1987;Charzynska et al., 1989;Comai et al., 1989;Turley and Trelease, 1990;Zhang et al., 1994). Thus, glyoxysomal enzymes are present at severa1 stages of the plant life cycle.Although genes encoding the key glyoxylate-cycle enzymes are activated in developing pollen and in maturing seed, the pathway's precise metabolic role(s) at these developmental stages is not known. Understanding the mechanisms that induce these...

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Analysis of the cucumber malate synthase gene promoter by transient expression and gel retardation assays

Abstract: SummaryRecently it has been demonstrated that the singlecopy malate eynthase (MS) and ieocitreta lyase (ICL) genes from cucumber are regulated by nutritional

Cited by 33 publications

References 27 publications

Activation of the Promoters of Arabidopsis Genes for the Branched-Chain α-Keto Acid Dehydrogenase Complex in Transgenic Tobacco BY-2 Cells under Sugar Starvation

Activation of the Promoters of Arabidopsis Genes for the Branched-Chain α-Keto Acid Dehydrogenase Complex in Transgenic Tobacco BY-2 Cells under Sugar Starvation

Molecular cloning and structural analysis of the cucumber (Cucumis sativus L.) phosphoenolpyruvate carboxykinase (CsPCK) gene

DNA Sequences That Activate Isocitrate Lyase Gene Expression during Late Embryogenesis and during Postgerminative Growth

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