Screening of Wound-Responsive Genes Identifies an Immediate-Early Expressed Gene Encoding a Highly Charged Protein in Mechanically Wounded Tobacco Plants

Hara, Kojiro; Yagi, Mitsue; Koizumi, Nozomu; Kusano, Tomonobu; Sano, Hiroshi

doi:10.1093/pcp/41.6.684

Cited by 47 publications

(43 citation statements)

References 32 publications

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“…Isolation of NtDRM1 cDNA-Initially, a DNA fragment encoding NtDRM1 was obtained by differential display of a cDNA population derived from wounded tobacco leaves (18). A full-length 2.4-kb cDNA of NtDRM1 was isolated by colony hybridization of a tobacco (N. tabacum) cDNA library using the NtDRM1 fragment as a probe and sequenced for both strands using a Big Dye terminator sequencing kit (Applied Biosystems, Foster City, CA).…”

Section: Methodsmentioning

confidence: 99%

Preferential de Novo Methylation of Cytosine Residues in Non-CpG Sequences by a Domains Rearranged DNA Methyltransferase from Tobacco Plants

Wada

Ohya

Yamaguchi

et al. 2003

Journal of Biological Chemistry

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In plant DNA, cytosines in symmetric CpG and CpNpG (N is A, T, or C) are thought to be methylated by DNA methyltransferases, MET1 and CMT3, respectively. Cytosines in asymmetric CpNpN are also methylated, and genetic analysis has suggested the responsible enzyme to be domains rearranged methyltransferase (DRM). We cloned a tobacco cDNA, encoding a novel protein consisting of 608 amino acids, that resembled DRMs found in maize and Arabidopsis and designated this as Nt-DRM1. The protein could be shown to be localized exclusively in the nucleus and exhibit methylation activity toward unmethylated synthetic as well as native DNA samples upon expression in Sf9 insect cells. It also methylated hemimethylated DNA, but the activity was lower than that for unmethylated substrates. Methylation mapping of a 962-bp DNA, treated with NtDRM1 in vitro, directly demonstrated methylation of ϳ70% of the cytosines in methylatable CpNpN and CpNpG sequences but only 10% in CpG. Further analyses indicated that the enzyme apparently non-selectively methylates any cytosines except in CpG, regardless of the adjacent nucleotide at both 5 and 3 ends. Transcripts of NtDRM1 ubiquitously accumulated in all tissues and during the cell cycle in tobacco cultured BY2 cells. These results indicate that NtDRM1 is a de novo cytosine methyltransfease, which actively excludes CpG substrate.The most commonly modified base in DNA among the eukaryotes through animals and plants is 5-methylcytosine (m 5 C).

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Section: Methodsmentioning

confidence: 99%

Preferential de Novo Methylation of Cytosine Residues in Non-CpG Sequences by a Domains Rearranged DNA Methyltransferase from Tobacco Plants

Wada

Ohya

Yamaguchi

et al. 2003

Journal of Biological Chemistry

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“…The searches revealed a high degree of sequence identity between GmMIPS1 and MIPS genes from plants and other organisms. The highest scoring match was to the tobacco MIPS sequence (Hara et al, 2000), which showed 80.6% identity to GmMIPS1 at the nucleotide level and 92.2% identity at the amino acid. Sequence identity to yeast was 40.2% at the nucleotide level and 41.5% at the amino acid level (Johnson and Henry, 1989), further confirming the high degree of conservation among MIPS sequences from different sources.…”

Section: Isolation and Analysis Of A Mips Cdnamentioning

confidence: 99%

“…A sequence similar to the yeast MIPS gene was identified in Spirodela polyrrhiza, an aquatic angiosperm (Smart and Fleming, 1993). MIPS sequences have been reported from Arabidopsis (Johnson, 1994), Citrus paradisii (Abu-Abied and Holland, 1994), common ice plant (Ishitani et al, 1996), and tobacco (Hara et al, 2000), and are highly conserved at the nucleotide level.…”

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confidence: 99%

Expression of d-myo-Inositol-3-Phosphate Synthase in Soybean. Implications for Phytic Acid Biosynthesis

2001

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Phytic acid, a phosphorylated derivative of myo-inositol, functions as the major storage form of phosphorus in plant seeds. Myo-inositol phosphates, including phytic acid, play diverse roles in plants as signal transduction molecules, osmoprotectants, and cell wall constituents. d-myo-inositol-3-phosphate synthase (MIPS EC 5.5.1.4) catalyzes the first step in de novo synthesis of myo-inositol. A soybean (Glycine max) MIPS cDNA (GmMIPS1) was isolated by reverse transcriptase-PCR using consensus primers designed from highly conserved regions in other plant MIPS sequences. Southern-blot analysis and database searches indicated the presence of at least four MIPS genes in the soybean genome. Northern-blot and immunoblot analyses indicated higher MIPS expression and accumulation in immature seeds than in other soybean tissues. MIPS was expressed early in the cotyledonary stage of seed development. The GmMIPS1 expression pattern suggested that it encodes a MIPS isoform that functions in seeds to generate d-myo-inositol-3-phosphate as a substrate for phytic acid biosynthesis.

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“…Treated leaf samples were harvested at appropriate time points and immediately frozen in liquid nitrogen before further analyses. The fluorescence differential display was performed essentially as described (Hara et al 2000), except that samples were fractionated by agarose gel electrophoresis.…”

mentioning

confidence: 99%

A stress-responsive multifunctional protein involved in .BETA.-oxidation in tobacco plants

Ohya

Ogata

Nakamura

et al. 2008

Plant Biotechnology

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Biotic and abiotic-induced wounding is one of the severest stresses that plants suffer throughout the growing period. Upon injury, plants rapidly activates a set of genes, which encode diverse proteins to cope with damages. Screening such genes that were transcriptionally activated within 30 min after mechanical wounding in tobacco plants, we identified a particular clone encoding a multfunctional protein, and designated as NtMFP (Nicotiana tabacum multifunctional protein). The deduced polypeptide is constituted of 668 amino acids with an apparent molecular mass of 72.3 kDa, and bacterially expressed protein exhibited a clear b-oxidation activity. Fusion proteins with GFP were observed in cytosol, when expressed in onion epidermal cell layers. In addition to wounding, NtMFP transcripts were induced by tobacco mosaic virus infection, and by jasmonic acid treatments. When NtMFP was suppressed by the RNAi, transgenic tobacco showed dwarfism, early senescence and reduced expresstion of jasmonic acid-responsive genes. Multifunctional protein is generally considered to catalyze multiple steps of the fatty acid b-oxidation. It is also proposed to be involved in the b-oxidation step of jasmonic acid biosynthesis. The present results suggest the possibility that NtMFP commonly functions not only in fatty acid catabolism but also in jasmonic acid biosynthesis pathway.

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Screening of Wound-Responsive Genes Identifies an Immediate-Early Expressed Gene Encoding a Highly Charged Protein in Mechanically Wounded Tobacco Plants

Cited by 47 publications

References 32 publications

Preferential de Novo Methylation of Cytosine Residues in Non-CpG Sequences by a Domains Rearranged DNA Methyltransferase from Tobacco Plants

Preferential de Novo Methylation of Cytosine Residues in Non-CpG Sequences by a Domains Rearranged DNA Methyltransferase from Tobacco Plants

Expression of d-myo-Inositol-3-Phosphate Synthase in Soybean. Implications for Phytic Acid Biosynthesis

A stress-responsive multifunctional protein involved in .BETA.-oxidation in tobacco plants

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