Cloning and characterization of a Nicotiana tabacum methylputrescine oxidase transcript

Heim, W; Sykes, Katie A.; Hildreth, Sherry B.; Sun, Jian; Lu, Rong-He; Jelesko, John G.

doi:10.1016/j.phytochem.2006.11.003

Cited by 94 publications

(73 citation statements)

References 31 publications

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“…S3). These expression patterns of NtMATE1/2 are very similar to those of nicotine biosynthetic genes (Hibi et al, 1994;Shoji et al, 2000aShoji et al, , 2002Reed and, Jelesko, 2004;Cane et al, 2005;Heim et al, 2007;Katoh et al, 2007).…”

Section: Expression Patternssupporting

confidence: 63%

“…Nicotine biosynthesis is positively regulated by the jasmonate-signaling cascade involving the COI1 F-box protein and JAZ repressors (Paschold et al, 2007;Shoji et al, 2008) and by the NIC regulatory loci that specifically control the gene expression of all enzymes known to be involved in the biosynthesis (Legg, 1984;Hibi et al, 1994;Reed and, Jelesko, 2004;Cane et al, 2005;Heim et al, 2007;Katoh et al, 2007). In flavonoid biosynthesis, regulatory genes coordinately regulate not only enzyme genes but also transporter genes responsible for intracellular transport of the metabolites (Koes et al, 2005).…”

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confidence: 99%

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Multidrug and Toxic Compound Extrusion-Type Transporters Implicated in Vacuolar Sequestration of Nicotine in Tobacco Roots

et al. 2008

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Nicotine is a major alkaloid accumulating in the vacuole of tobacco (Nicotiana tabacum), but the transporters involved in the vacuolar sequestration are not known. We here report that tobacco genes (NtMATE1 and NtMATE2) encoding transporters of the multidrug and toxic compound extrusion (MATE) family are coordinately regulated with structural genes for nicotine biosynthesis in the root, with respect to spatial expression patterns, regulation by NIC regulatory loci, and induction by methyl jasmonate. Subcellular fractionation, immunogold electron microscopy, and expression of a green fluorescent protein fusion protein all suggested that these transporters are localized to the vacuolar membrane. Reduced expression of the transporters rendered tobacco plants more sensitive to the application of nicotine. In contrast, overexpression of NtMATE1 in cultured tobacco cells induced strong acidification of the cytoplasm after jasmonate elicitation or after the addition of nicotine under nonelicited conditions. Expression of NtMATE1 in yeast (Saccharomyces cerevisiae) cells compromised the accumulation of exogenously supplied nicotine into the yeast cells. The results imply that these MATE-type proteins transport tobacco alkaloids from the cytosol into the vacuole in exchange for protons in alkaloid-synthesizing root cells.

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Section: Expression Patternssupporting

confidence: 63%

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confidence: 99%

Multidrug and Toxic Compound Extrusion-Type Transporters Implicated in Vacuolar Sequestration of Nicotine in Tobacco Roots

et al. 2008

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“…Nicotine is composed of a pyridine ring and a pyrrolidine ring (see Supplemental Figure 1 online); ornithine decarboxylase (ODC; Imanishi et al, 1998), putrescine N-methyltransferase (PMT; Hibi et al, 1994), and N-methylputrescine oxidase (MPO; Heim et al, 2007;Katoh et al, 2007) are involved in the formation of the pyrrolidine ring, whereas the pyridine ring uses enzymes involved in the early steps of NAD biosynthesis, such as aspartate oxidase (AO), quinolinic acid synthase (QS), and quinolinic acid phosphoribosyl transferase (QPT) (Sinclair et al, 2000;Katoh et al, 2006). A PIP family oxidoreductase, A622, is also required for the biosynthesis of tobacco alkaloids, possibly to form a nicotinic acid-derived precursor (Deboer et al, 2009;Kajikawa et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Clustered Transcription Factor Genes Regulate Nicotine Biosynthesis in Tobacco

2010

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Tobacco (Nicotiana tabacum) synthesizes nicotine and related pyridine alkaloids in the root, and their synthesis increases upon herbivory on the leaf via a jasmonate-mediated signaling cascade. Regulatory NIC loci that positively regulate nicotine biosynthesis have been genetically identified, and their mutant alleles have been used to breed low-nicotine tobacco varieties. Here, we report that the NIC2 locus, originally called locus B, comprises clustered transcription factor genes of an ethylene response factor (ERF) subfamily; in the nic2 mutant, at least seven ERF genes are deleted altogether. Overexpression, suppression, and dominant repression experiments using transgenic tobacco roots showed both functional redundancy and divergence among the NIC2-locus ERF genes. These transcription factors recognized a GCC-box element in the promoter of a nicotine pathway gene and specifically activated all known structural genes in the pathway. The NIC2-locus ERF genes are expressed in the root and upregulated by jasmonate with kinetics that are distinct among the members. Thus, gene duplication events generated a cluster of highly homologous transcription factor genes with transcriptional and functional diversity. The NIC2-locus ERFs are close homologs of ORCA3, a jasmonate-responsive transcriptional activator of indole alkaloid biosynthesis in Catharanthus roseus, indicating that the NIC2/ORCA3 ERF subfamily was recruited independently to regulate jasmonate-inducible secondary metabolism in distinct plant lineages.

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“…Putrescine is methylated by putrescine methyltransferase (PMT) to yield N-methylputrescine, which is subsequently converted into 4-methylaminobutanal by N-methylputrescine oxidase (Heim et al, 2007;Katoh et al, 2007;Biastoff et al, 2009). 4-Methylaminobutanal is postulated to undergo spontaneous cyclization to form an N-methyl-D 1 -pyrrolinium cation, which is ultimately converted to tropinone through an unknown mechanism (O'Connor, 2010;Jirschitzka et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

A Root-Expressed l-Phenylalanine:4-Hydroxyphenylpyruvate Aminotransferase Is Required for Tropane Alkaloid Biosynthesis in Atropa belladonna

et al. 2014

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The tropane alkaloids, hyoscyamine and scopolamine, are medicinal compounds that are the active components of several therapeutics. Hyoscyamine and scopolamine are synthesized in the roots of specific genera of the Solanaceae in a multistep pathway that is only partially elucidated. To facilitate greater understanding of tropane alkaloid biosynthesis, a de novo transcriptome assembly was developed for Deadly Nightshade (Atropa belladonna). Littorine is a key intermediate in hyoscyamine and scopolamine biosynthesis that is produced by the condensation of tropine and phenyllactic acid. Phenyllactic acid is derived from phenylalanine via its transamination to phenylpyruvate, and mining of the transcriptome identified a phylogenetically distinct aromatic amino acid aminotransferase (ArAT), designated Ab-ArAT4, that is coexpressed with known tropane alkaloid biosynthesis genes in the roots of A. belladonna. Silencing of Ab-ArAT4 disrupted synthesis of hyoscyamine and scopolamine through reduction of phenyllactic acid levels. Recombinant Ab-ArAT4 preferentially catalyzes the first step in phenyllactic acid synthesis, the transamination of phenylalanine to phenylpyruvate. However, rather than utilizing the typical keto-acid cosubstrates, 2-oxoglutarate, pyruvate, and oxaloacetate, Ab-ArAT4 possesses strong substrate preference and highest activity with the aromatic keto-acid, 4-hydroxyphenylpyruvate. Thus, Ab-ArAT4 operates at the interface between primary and specialized metabolism, contributing to both tropane alkaloid biosynthesis and the direct conversion of phenylalanine to tyrosine.

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Cloning and characterization of a Nicotiana tabacum methylputrescine oxidase transcript

Cited by 94 publications

References 31 publications

Multidrug and Toxic Compound Extrusion-Type Transporters Implicated in Vacuolar Sequestration of Nicotine in Tobacco Roots

Multidrug and Toxic Compound Extrusion-Type Transporters Implicated in Vacuolar Sequestration of Nicotine in Tobacco Roots

Clustered Transcription Factor Genes Regulate Nicotine Biosynthesis in Tobacco

A Root-Expressed l-Phenylalanine:4-Hydroxyphenylpyruvate Aminotransferase Is Required for Tropane Alkaloid Biosynthesis in Atropa belladonna

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