Regulation of a Sesquiterpene Cyclase in Cellulase-Treated Tobacco Cell Suspension Cultures

Vögeli, Urs; Chappell, Joseph

doi:10.1104/pp.94.4.1860

Cited by 43 publications

(25 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, the level of cyclase mRNA translational activity in elicitor-treated cells was similar to that previously described (37). The cyclase translational activity was substantially reduced when elicitor-treated cell cultures were exposed to the calmodulin antagonists, 15 to 72% depending on antagonist and concentration used.…”

Section: Calcium/calmodulin Antagonists Suppresssupporting

confidence: 85%

“…Immunoblot analysis of proteins separated by SDS-PAGE and specific immunoprecipitation of the radioactive cyclase and PAL proteins produced in vivo or in vitro have been described previously (37). The PAL antibodies were kindly provided by Dr. Lee Hadwiger (Washington State University, Pullman, WA).…”

Section: Immunological Techniquesmentioning

confidence: 99%

“…Previous work correlated the induction of sesquiterpenoid accumulation with an induction of a sesquiterpene cyclase, 5-epi-aristolochene synthase (34,36). Regulation of the cyclase enzyme activity was recently demonstrated to be controlled primarily by the transcription rate of the respective gene (37) KY 14 were maintained in Murashige-Skoog medium as previously described (8). Cell cultures in their rapid phase of growth, corresponding to 3 d after subculturing, were used for all the experiments presented.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Inhibition of Phytoalexin Biosynthesis in Elicitor-Treated Tobacco Cell-Suspension Cultures by Calcium/Calmodulin Antagonists

Vögeli¹,

Vögeli‐Lange²,

Chappell³

1992

Plant Physiol.

Self Cite

View full text Add to dashboard Cite

A role for calcium/calcium-binding proteins in a mechanism of signaling elicitor-inducible phytoalexin biosynthesis was investigated. Two classes of calcium/calmodulin antagonists, phenothiazines and naphthalenesulfonamides, inhibited sesquiterpene phytoalexin accumulation in tobacco (Nicotiana tabacum) cellsuspension cultures when added 1 h before elicitor. The antagonists also inhibited the induction of sesquiterpene cyclase enzyme activity, a key regulatory enzyme for sesquiterpene biosynthesis. The antagonists suppressed the induction of sesquiterpene cyclase only if added before or simultaneously with elicitor. Additionally, the antagonists inhibited (a) accumulation of the cyclase protein as measured in immunoblots; (b) the in vivo synthesis rate of the cyclase protein, measured as the incorporation of [35S]methionine into immunoprecipitable cyclase protein; and (c) the cyclase mRNA translational activity, measured as the incorporation of [35S]-methionine into immunoprecipitable cyclase protein synthesized by in vitro translation of RNA isolated from antagonist-treated, elicitor-induced cells. In contrast, elicitor-inducible phenylalanine ammonia lyase enzyme activity, the level of the enzyme protein, the in vivo synthesis rate, and the mRNA translational activity were not affected by any of the antagonist treatments. Uptake and incorporation of [35S]methionine into total cellular proteins and total in vitro translation products were also not indiscriminately altered by the antagonist treatments. The current results suggest that calcium and/or calmodulin-like proteins may be elements of a signal transduction pathway mediating elicitor-induced accumulation of phytoalexins in tobacco.Plants can respond to pathogen challenge by the induction of a number of biochemical responses (3), including phytoalexin accumulation (11, 13), synthesis and secretion of hydrolytic enzymes (4), and the putative reinforcement of their cell walls with lignin and/or structural glycoproteins (32).

show abstract

Section: Calcium/calmodulin Antagonists Suppresssupporting

confidence: 85%

Section: Immunological Techniquesmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Inhibition of Phytoalexin Biosynthesis in Elicitor-Treated Tobacco Cell-Suspension Cultures by Calcium/Calmodulin Antagonists

Vögeli¹,

Vögeli‐Lange²,

Chappell³

1992

Plant Physiol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Previous studies have demonstrated that inducible monoterpene (Steele et al, 1998), sesquiterpene (Vö geli and Chappell, 1990;Facchini and Chappell, 1992) and diterpene (Lois and West, 1990) biosynthesis are regulated at the level of transcription. The present studies are the first to indicate that the developmental regulation of monoterpene biosynthesis in secretory trichomes also resides largely at the level of gene expression.…”

Section: Transcriptional and Translational Regulation Of Monoterpene mentioning

confidence: 99%

Developmental Regulation of Monoterpene Biosynthesis in the Glandular Trichomes of Peppermint

Gershenzon²,

2000

View full text Add to dashboard Cite

show abstract

“…The induction of EAS enzyme activity has been correlated with the absolute amount and the de novo synthesis of the enzyme protein (12). Changes in the de novo synthesis rate of the EAS protein were also correlated with changes in the EAS mRNA translational activity.…”

Section: Introductionmentioning

confidence: 95%

Gene family for an elicitor-induced sesquiterpene cyclase in tobacco.

Facchini

Chappell

1992

Proc. Natl. Acad. Sci. U.S.A.

237

174

View full text Add to dashboard Cite

The initial step in the conversion of the isoprenoid intermediate farnesyl diphosphate to the sesquiterpenoid phytoalexin capsidiol in elicitor-treated tobacco tissues is catalyzed by an inducible sesquiterpene cyclase [5-epiaristolochene synthase (EAS)]. Two independent cDNA clones (cEAS1 and cEAS2) encoding EAS were isolated from an elicitor-induced tobacco cDNA library by differential hybridization and subsequently were characterized by hybrid selection-in vitro translation. Insertion of cEAS1, a partial cDNA clone encoding 175 C-terminal amino acids, into an Escherchia coli expression vector resulted in accumulation of a fusion protein immunodetectable with EAS-specific polyclonal antibodies. The cDNA clones were used to isolate two full-length EAS genes that mapped 5 kilobases (kb) apart on one 15-kb genomic clone. The nucleotide sequences of the structural gene components were identical from 388 base pairs (bp) upstream of the transcription initiation site to 40 bp downstream of the translation termination codon, suggesting a relatively recent duplication event. The genes consist of 1479-bp open reading frames, each containing five introns and specifying 56,828-Da proteins. The N-terminal amino acid sequence deduced from the genomic clones was identical to the first 16 amino acids of the EAS protein identifuible by Edman degradation. RNA blot hybridization with cEAS1 demonstrated a mRNA induction time course consistent with the induction of the EAS mRNA translational activity with maximum levels 4-6 h after elicitation. EAS mRNA was not detected in control cells. DNA blot-hybridization analysis of genomic DNA revealed a copy number of '12-15 for EAS-like genes in the tetraploid tobacco genome. The conservation of a putative allelic prenyl diphosphate binding motif is also discussed.Sesquiterpenoids are a structurally diverse class of isoprenoids found in plants, some fungi, and bacteria, but not in vertebrates, which have important implications in plantplant (1), plant-insect (2, 3), and plant-pathogen (4, 5) interactions. Despite an extensive appreciation for the structure-function relationships of plant sesquiterpenoids, the sesquiterpenoid biosynthetic pathway and its regulation are not well understood (4, 6). Although many enzymes of general isoprenoid biosynthesis have been measured in plants, few enzymes suspected of being rate-limiting have been identified (4, 6). Moreover, little is known about the mechanisms regulating the activity and absolute levels of plant isoprenoid biosynthetic enzymes with the exception of a diterpene cyclase (7). This is in contrast to the extensive molecular analysis of regulatory mechanisms controlling isoprenoid metabolism in mammalian systems (8).Tobacco cell suspension cultures respond to treatment with elicitors such as fungal cell wall hydrolysates or cellulase by the de novo synthesis and secretion of antibiotic sesquiterpenoids, primarily the phytoalexin capsidiol (9, 10). Central to capsidiol biosynthesis is the induction of a sesquiterpene cyclase [5-epi...

show abstract

Regulation of a Sesquiterpene Cyclase in Cellulase-Treated Tobacco Cell Suspension Cultures

Cited by 43 publications

References 16 publications

Inhibition of Phytoalexin Biosynthesis in Elicitor-Treated Tobacco Cell-Suspension Cultures by Calcium/Calmodulin Antagonists

Inhibition of Phytoalexin Biosynthesis in Elicitor-Treated Tobacco Cell-Suspension Cultures by Calcium/Calmodulin Antagonists

Developmental Regulation of Monoterpene Biosynthesis in the Glandular Trichomes of Peppermint

Gene family for an elicitor-induced sesquiterpene cyclase in tobacco.

Contact Info

Product

Resources

About