Nucleotide sequence of the tryptophan decarboxylase gene of Catharanthus roseus and expression of tdc-gusA gene fusions in Nicotiana tabacum

Goddijn, Oscar J. M.; Lohman, Frans P.; Kam, Rolf J. de; hilperoort, Rob A.; Hoge, J. H. C.

doi:10.1007/bf00391016

Cited by 53 publications

(35 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At present, cDNA and genomic clones for the TIA biosynthetic genes Tdc (De Luca et al 1989;Goddijn et al 1994), Str (McKnight et al 1990;Pasquali et al 1992Pasquali et al , 1999 and Cpr (Meijer et al 1993b;Lopes-Cardoso et al 1997) have been isolated from C. roseus. In cell suspensions, the Tdc, Str and Cpr genes are coordinately induced by fungal elicitors (Pasquali et al 1992;Roewer et al 1992;Meijer et al 1993b), and repressed by auxin Pasquali et al 1992;Meijer 1993).…”

Section: Introductionmentioning

confidence: 98%

“…Constitutive expression of the Tdc coding region under control of the strong cauli¯ower mosaic virus (CaMV) 35S promoter enhanced TDC enzyme activity up to tenfold in transgenic C. roseus callus (Goddijn et al 1995), indicating that TDC enzyme levels are determined mainly by the expression level of the Tdc gene. Tdc promoter sequences were shown to confer reporter gene activity in leaves, stems and roots of transgenic tobacco plants (Goddijn et al 1994). Progressive 5¢ deletions gradually decreased promoter strength.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Nuclear factors GT-1 and 3AF1 interact with multiple sequences within the promoter of the Tdc gene from Madagascar periwinkle: GT-1 is involved in UV light-induced expression

Ouwerkerk¹,

Trimborn²,

Hilliou³

et al. 1999

Mol Gen Genet

View full text Add to dashboard Cite

Plant secondary metabolites of the terpenoid indole alkaloid (TIA) class comprise several compounds with pharmaceutical applications. A key step in the TIA biosynthetic pathway is catalysed by the enzyme tryptophan decarboxylase (TDC), which channels the primary metabolite tryptophan into TIA metabolism. In Catharanthus roseus (Madagascar periwinkle), the Tdc gene is expressed throughout plant development. Moreover, Tdc gene expression is induced by external stress signals, such as fungal elicitor and UV light. In a previous study of Tdc promoter architecture in transgenic tobacco it was shown that the -538 to -112 region is a quantitative determinant for the expression level in different plant organs. Within this sequence one particular region (-160 to -99) was identified as the major contributor to basal expression and another region (-99 to -37) was shown to be required for induction by fungal elicitor. Here, the in vitro binding of nuclear factors to the -572 to -37 region is described. In extracts from tobacco and C. roseus, two binding activities were detected that could be identified as the previously described nuclear factors GT-1 and 3AF1, based on their mobility and binding characteristics. Both factors appeared to interact with multiple regions in the Tdc promoter. Mutagenesis of GT-1 binding sites in the Tdc promoter did not affect the basal or elicitor-induced expression levels. However, induction of the Tdc promoter constructs by UV light was significantly lower, thereby demonstrating a functional role for GT-1 in the induction of Tdc expression by UV light.

show abstract

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

Nuclear factors GT-1 and 3AF1 interact with multiple sequences within the promoter of the Tdc gene from Madagascar periwinkle: GT-1 is involved in UV light-induced expression

Ouwerkerk¹,

Trimborn²,

Hilliou³

et al. 1999

Mol Gen Genet

View full text Add to dashboard Cite

show abstract

“…The indole moiety of ajmalicine is derived from tryptamine, which is directly synthesized from tryptophan by tryptophan decarboxylase (TDC, EC 4.1.1.28) (Goddijn et al 1994). Tryptophan decarboxylase plays a key role in TIAs biosynthesis by linking primary and secondary metabolism (Valletta et al 2010).…”

Section: Introductionmentioning

confidence: 99%

Tryptophan decarboxylase plays an important role in ajmalicine biosynthesis in Rauvolfia verticillata

Liu

Chen

et al. 2012

Planta

View full text Add to dashboard Cite

Tryptophan decarboxylase (TDC) converts tryptophan into tryptamine that is the indole moiety of ajmalicine. The full-length cDNA of Rauvolfia verticillata (RvTDC) was 1,772 bps that contained a 1,500-bp ORF encoding a 499-amino-acid polypeptide. Recombinant 55.5 kDa RvTDC converted tryptophan into tryptamine. The K (m) of RvTDC for tryptophan was 2.89 mM, higher than those reported in other TIAs-producing plants. It demonstrated that RvTDC had lower affinity to tryptophan than other plant TDCs. The K (m) of RvTDC was also much higher than that of strictosidine synthase and strictosidine glucosidase in Rauvolfia. This suggested that TDC might be the committed-step enzyme involved in ajmalicine biosynthesis in R. verticillata. The expression of RvTDC was slightly upregulated by MeJA; the five MEP pathway genes and SGD showed no positive response to MeJA; and STR was sharply downregulated by MeJA. MeJA-treated hairy roots produced higher level of ajmalicine (0.270 mg g(-1) DW) than the EtOH control (0.183 mg g(-1) DW). Highest RvTDC expression level was detected in hairy root, about respectively 11, 19, 65, and 109-fold higher than in bark, young leaf, old leaf, and root. Highest ajmalicine content was also found in hairy root (0.249 mg g(-1) DW) followed by in bark (0.161 mg g(-1) DW) and young leaf (0.130 mg g(-1) DW), and least in root (0.014 mg g(-1) DW). Generally, the expression level of RvTDC was positively consistent with the accumulation of ajmalicine. Therefore, it could be deduced that TDC might be the key enzyme involved in ajmalicine biosynthesis in Rauvolfia.

show abstract

“…Genomic library construction and screening C. roseus total DNA, partially digested with Sau3AI, was cloned in kGEM11 (Promega) BamHI arms (Goddijn et al 1994). Library screening was performed as described in Memelink et al (1994).…”

Section: Methodsmentioning

confidence: 99%

A promoter region that controls basal and elicitor-inducible expression levels of the NADPH:cytochrome P450 reductase gene (Cpr ) from Catharanthus roseus binds nuclear factor GT-1

Cardoso¹,

Meijer²,

Rueb³

et al. 1997

Mol Gen Genet

View full text Add to dashboard Cite

NADPH:cytochrome P450 reductase (CPR) is essential for the activation of cytochrome P450 enzymes, which are involved in a wide variety of metabolic pathways in plants, including those related to defence responses. In the subtropical plant Catharanthus roseus several cytochrome P450 enzymes operate in the biosynthesis of defence-related terpenoid indole alkaloids (TIAs). In agreement with the importance of CPR in defence, Cpr mRNA levels in C. roseus were found to be enhanced by fungal elicitor preparations that also induce TIA biosynthesis and P450 gene expression. Here we describe the isolation of a C. roseus genomic DNA clone covering the 5¢ part of the Cpr gene and 1.6-kb of upstream sequences. Mapping of the transcription start site showed the untranslated leader sequence is approximately 280 bp long. To study the control of gene expression by the Cpr promoter, transcriptional fusions between Cpr promoter fragments and the gusA reporter gene were generated and their expression was analyzed in stably transformed tobacco plants. The Cpr promoter fragment extending from A1510 to A8, with respect to the ATG start codon, conferred basal and elicitor-inducible expression on the gusA reporter gene, strongly indicating that the Cpr gene of C. roseus is indeed controlled by this promoter region. Progressive deletion from the 5¢ end of the promoter to position A632 had little eect on gusA expression. However, deletion to position A366 resulted in a complete loss of basal activity and largely eliminated elicitor-induced expression, indicating that the region from A632 to A366 contains the main transcription-enhancing cis-regulatory sequences. Electrophoretic mobility shift assays with tobacco nuclear extracts showed that binding sites for nuclear factor GT-1 are redundant in the Cpr promoter, but absent from the downstream part of the leader sequence. The presence of strong GT-1 binding sites in the main enhancer region (A632 to A366), is suggestive of a functional role for this factor in basal expression and elicitor responsiveness of the Cpr promoter.

show abstract

Nucleotide sequence of the tryptophan decarboxylase gene of Catharanthus roseus and expression of tdc-gusA gene fusions in Nicotiana tabacum

Cited by 53 publications

References 22 publications

Nuclear factors GT-1 and 3AF1 interact with multiple sequences within the promoter of the Tdc gene from Madagascar periwinkle: GT-1 is involved in UV light-induced expression

Nuclear factors GT-1 and 3AF1 interact with multiple sequences within the promoter of the Tdc gene from Madagascar periwinkle: GT-1 is involved in UV light-induced expression

Tryptophan decarboxylase plays an important role in ajmalicine biosynthesis in Rauvolfia verticillata

A promoter region that controls basal and elicitor-inducible expression levels of the NADPH:cytochrome P450 reductase gene (Cpr ) from Catharanthus roseus binds nuclear factor GT-1

Contact Info

Product

Resources

About