Research Interests: Genetics, Plant Biology, Biology, Cell Biology, Medicine, and 15 moreHumans, Sequence alignment, Proteins, Plasmids, Arabidopsis, Amino Acid Sequence, Base Sequence, Recombinant Proteins, Cytoplasm, PLANT PROTEINS, Protein Transport, Biochemistry and cell biology, Dimerization, Intracellular, and Molecular Sequence Data
Jasmonate (JA) and gibberellins (GAs) exert antagonistic effects on plant growth and development in response to environmental and endogenous stimuli. Although the crosstalk between JA and GA has been elucidated, the role of JA in GA... more
Jasmonate (JA) and gibberellins (GAs) exert antagonistic effects on plant growth and development in response to environmental and endogenous stimuli. Although the crosstalk between JA and GA has been elucidated, the role of JA in GA biosynthesis remains unclear. Therefore, in this study, we investigated the mechanism underlying JA-mediated regulation of endogenous GA levels in Arabidopsis (Arabidopsis thaliana). Transient and electrophoretic mobility shift assays showed that transcription factor MYC2 regulates GA inactivation genes. Using transgenic plants, we further evaluated the contribution of MYC2 in regulating GA inactivation genes. JA treatment increased DELLA accumulation but did not inhibit DELLA protein degradation. Additionally, JA treatment decreased bioactive GA content, including GA4, significantly decreased the expression of GA biosynthesis genes, including ent-kaurene synthase (AtKS), GA 3β-hydroxylase (AtGA3ox1), and AtGA3ox2, and increased the expression of GA inactivation genes, including GA 2 oxidase (AtGA2ox4), AtGA2ox7, and AtGA2ox9. Conversely, JA treatment did not significantly affect gene expression in the myc2 myc3 myc4 triple mutant, demonstrating the MYC2–4-dependent effects of JA in GA biosynthesis. Additionally, JA post-transcriptionally regulated AtGA3ox1 expression. We identified microRNA miR5998 as an AtGA3ox1-associated miRNA; its overexpression inhibited plant growth by suppressing AtGA3ox1 expression. Overall, our findings indicate that JA treatment inhibits endogenous GA levels and plant growth by decreasing the expression of GA biosynthesis genes and increasing the expression of GA inactivation genes via miR5998 and MYC2 activities.
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KEY MESSAGE The GAF1 transcription factor is shown to bind to the promoter of the Arabidopsis GA-biosynthetic enzyme GA20ox1 and, in association with DELLA protein, promotes GA20ox1 expression, thereby contributing to its feedback... more
KEY MESSAGE The GAF1 transcription factor is shown to bind to the promoter of the Arabidopsis GA-biosynthetic enzyme GA20ox1 and, in association with DELLA protein, promotes GA20ox1 expression, thereby contributing to its feedback regulation and tissue specificity. Gibberellins (GAs) are phytohormones that promote plant growth and development, including germination, elongation, flowering, and floral development. Homeostasis of endogenous GA levels is controlled by GA feedback regulation. DELLAs are negative regulators of GA signaling that are rapidly degraded in the presence of GAs. DELLAs regulate several target genes, including AtGA20ox2 and AtGA3ox1, encoding the GA-biosynthetic enzymes GA 20-oxidase and GA 3-oxidase, respectively. Previous studies have identified GAI-ASSOCIATED FACTOR 1 (GAF1) as a DELLA interactor, with which DELLAs act as transcriptional coactivators; furthermore, AtGA20ox2, AtGA3ox1, and AtGID1b were identified as target genes of the DELLA-GAF1 complex. Among the five Arabidopsis GA20ox genes, AtGA20ox1 is the most highly expressed gene during vegetative growth; its expression is controlled by GA feedback regulation. Here, we investigated whether AtGA20ox1 is regulated by the DELLA-GAF1 complex. The electrophoretic mobility shift and transactivation assays showed that three GAF1-binding sites exist in the AtGA20ox1 promoter. Using transgenic plants, we further evaluated the contribution of the DELLA-GAF1 complex to GA feedback regulation and tissue-specific expression. Mutations in two GAF1-binding sites obliterated the negative feedback regulation and tissue-specific expression of AtGA20ox1 in transgenic plants. Thus, our results showed that GAF1-binding sites are involved in GA feedback regulation and tissue-specific expression of AtGA20ox1 in Arabidopsis, suggesting that the DELLA-GAF1 complex is involved in both processes.
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Our study focused on the functional analysis of RSG, a tobacco transcriptional activator with a bZIP domain. Expression of a dominant-negative form of RSG severely inhibited the process of cell elongation in stems and reduced the... more
Our study focused on the functional analysis of RSG, a tobacco transcriptional activator with a bZIP domain. Expression of a dominant-negative form of RSG severely inhibited the process of cell elongation in stems and reduced the endogenous amounts of GA 1, which is the major active gibberellin (GA) in tobacco in transgenic plants. To investigate the function of RSG in
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Gibberellins (GAs) are phytohormones that regulate many aspects of plant growth and development, including germination, elongation, flowering, and floral development. Negative feedback regulation contributes to homeostasis of the GA... more
Gibberellins (GAs) are phytohormones that regulate many aspects of plant growth and development, including germination, elongation, flowering, and floral development. Negative feedback regulation contributes to homeostasis of the GA level. DELLAs are negative regulators of GA signaling and are rapidly degraded in the presence of GAs. DELLAs regulate many target genes, including AtGA20ox2 in Arabidopsis (Arabidopsis thaliana), encoding the GA-biosynthetic enzyme GA 20-oxidase. As DELLAs do not have an apparent DNA-binding motif, transcription factors that act in association with DELLA are necessary for regulating the target genes. Previous studies have identified GAI-ASSOCIATED FACTOR1 (GAF1) as such a DELLA interactor, with which DELLAs act as coactivators, and AtGA20ox2 was identified as a target gene of the DELLA-GAF1 complex. In this study, electrophoretic mobility shift and chromatin immunoprecipitation assays showed that four GAF1-binding sites exist in the AtGA20ox2 promoter. Using transgenic plants, we further evaluated the contribution of the DELLA-GAF1 complex to GA feedback regulation. Mutations in four GAF1-binding sites abolished the negative feedback of AtGA20ox2 in transgenic plants. Our results showed that GAF1-binding sites are necessary for GA feedback regulation of AtGA20ox2, suggesting that the DELLA-GAF1 complex is a main component of the GA feedback regulation of AtGA20ox2.
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Research Interests: Genetics, Plant Biology, Biology, Calcium, Cell Biology, and 15 moreMedicine, Arabidopsis thaliana, Protein Kinases, Mutation, Arabidopsis, Kinase, Nicotiana Tabacum, Arginine, Plant cell, Calcium Binding Proteins, Structure activity Relationship, Protein Binding, PLANT PROTEINS, Biochemistry and cell biology, and Molecular Sequence Data
Research Interests: Genetics, Plant Biology, Biology, Cell Biology, Medicine, and 15 moreHumans, Sequence alignment, Proteins, Plasmids, Arabidopsis, Amino Acid Sequence, Base Sequence, Recombinant Proteins, Cytoplasm, PLANT PROTEINS, Protein Transport, Biochemistry and cell biology, Dimerization, Intracellular, and Molecular Sequence Data
Research Interests: Genetics, Plant Biology, Biology, Tobacco, Medicine, and 15 moreProteins, Green Fluorescent Protein, Gibberellin, Enzyme, Nicotiana Tabacum, Protein Degradation, Plant cell, Amino Acid Sequence, Protein Binding, Gibberellins, Cytoplasm, Transgenic plant, Biochemistry and cell biology, Molecular Sequence Data, and reverse transcriptase polymerase chain reaction
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Research Interests: Genetics, Plant Biology, Biology, Medicine, Arabidopsis, and 15 moreGibberellin, Phenotype, Enzyme, Molecular cloning, Elongation, Plant cell, Amino Acid Sequence, Base Sequence, Gibberellins, Full Length Movies, Cell Expansion, Biochemistry and cell biology, Cell Size, Molecular Sequence Data, and Leucine zipper
Jasmonate (JA) and gibberellins (GAs) exert antagonistic effects on plant growth and development in response to environmental and endogenous stimuli. Although the crosstalk between JA and GA has been elucidated, the role of JA in GA... more
Jasmonate (JA) and gibberellins (GAs) exert antagonistic effects on plant growth and development in response to environmental and endogenous stimuli. Although the crosstalk between JA and GA has been elucidated, the role of JA in GA biosynthesis remains unclear. Therefore, in this study, we investigated the mechanism underlying JA-mediated regulation of endogenous GA levels in Arabidopsis (Arabidopsis thaliana). Transient and electrophoretic mobility shift assays showed that transcription factor MYC2 regulates GA inactivation genes. Using transgenic plants, we further evaluated the contribution of MYC2 in regulating GA inactivation genes. JA treatment increased DELLA accumulation but did not inhibit DELLA protein degradation. Additionally, JA treatment decreased bioactive GA content, including GA4, significantly decreased the expression of GA biosynthesis genes, including ent-kaurene synthase (AtKS), GA 3β-hydroxylase (AtGA3ox1), and AtGA3ox2, and increased the expression of GA inac...
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Flowering is the developmental transition from the vegetative to the reproductive phase. FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1), and LEAFY (LFY) are floral integrators. These genes are repressed by... more
Flowering is the developmental transition from the vegetative to the reproductive phase. FLOWERING LOCUS T (FT), SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1), and LEAFY (LFY) are floral integrators. These genes are repressed by several floral repressors including EARLY FLOWERING3 (ELF3), SHORT VEGETATIVE PHASE (SVP), TEMPRANILLO1 (TEM1), and TEM2. Although gibberellin (GA) promotes flowering by activating the floral integrator genes, the exact molecular mechanism remains unclear. DELLAs are negative regulators in GA signaling and act as coactivators of the transcription factor GAI ASSOCIATED FACTOR 1 (GAF1). GAs convert the GAF1 complex from a transcriptional activator to a repressor. Here, we show that GAF1 functions in the GA-dependent flowering pathway by regulating FT and SOC1 expression in Arabidopsis thaliana. We identified four flowering repressors, ELF3, SVP, TEM1, and TEM2, as GAF1-target genes. In response to GAs, GAF1 forms a transcriptional repressor complex and prom...
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DELLA proteins act as negative regulators in gibberellin (GA) signal transduction. GA-induced DELLA degradation is a central regulatory system in GA signaling pathway. Intensive studies have revealed the degradation mechanism of DELLA and... more
DELLA proteins act as negative regulators in gibberellin (GA) signal transduction. GA-induced DELLA degradation is a central regulatory system in GA signaling pathway. Intensive studies have revealed the degradation mechanism of DELLA and the functions of DELLA as a transcriptional regulator. Meanwhile, recent studies suggest the existence of a DELLA-independent GA signaling pathway. In this review, we summarized the DELLA-independent GA signaling pathway together with the well-analyzed DELLA-dependent pathway.
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DELLA proteins play a central role in gibberellin (GA) signaling. GA triggers DELLA degradation via the ubiquitin-proteasome pathway, thereby promoting plant growth. An increase in cytosolic Ca2+ ([Ca2+]cyt) was previously observed after... more
DELLA proteins play a central role in gibberellin (GA) signaling. GA triggers DELLA degradation via the ubiquitin-proteasome pathway, thereby promoting plant growth. An increase in cytosolic Ca2+ ([Ca2+]cyt) was previously observed after several hours GA application. Recent studies also suggest the existence of a DELLA-independent GA response. However, the effect of DELLA on the GA-induced increase in [Ca2+]cyt remains unknown. This study re-examined the effects of GAs on [Ca2+]cyt using the Ca2+ sensor protein aequorin. [Ca2+]cyt increased within a few minutes of GA treatment, even in transgenic plants expressing a mutated degradation-resistant version of RGA and in della pentuple mutant plants. In addition, it was also revealed that Ca2+ is not involved in DELLA degradation. These results suggest that the GA-induced increase in [Ca2+]cyt occurs via a DELLA-independent pathway, providing important information on the GA signaling network.
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Protein kinases regulate diverse physiological processes. Because many kinases preserve inherent autophosphorylation capability, autophosphorylation appears to be one of the most important mechanisms for cellular signaling. However,... more
Protein kinases regulate diverse physiological processes. Because many kinases preserve inherent autophosphorylation capability, autophosphorylation appears to be one of the most important mechanisms for cellular signaling. However, physiological functions of autophosphorylation are still largely unknown, other than the self-activation by phosphorylation of activation loop in the catalytic domain. REPRESSION OF SHOOT GROWTH (RSG) is the transcription factor involved in gibberellin (GA) feedback regulation. The tobacco (Nicotiana tabacum) Ca(2+)-dependent protein kinase, NtCDPK1, phosphorylates RSG, resulting in the negative regulation of RSG. NtCDPK1 was previously shown to be autophosphorylated in a Ca(2+)-dependent manner. Here, we investigated the functional importance of autophosphorylation in NtCDPK1. Ser-6 and Thr-21 were identified as autophosphorylation sites of NtCDPK1. Autophosphorylation not only reduced the binding affinity of NtCDPK1 for RSG, but also inhibited the homo...
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Our study focused on the functional analysis of RSG, a tobacco transcriptional activator with a bZIP domain. Expression of a dominant-negative form of RSG severely inhibited the process of cell elongation in stems and reduced the... more
Our study focused on the functional analysis of RSG, a tobacco transcriptional activator with a bZIP domain. Expression of a dominant-negative form of RSG severely inhibited the process of cell elongation in stems and reduced the endogenous amounts of GA 1, which is the major active gibberellin (GA) in tobacco in transgenic plants. To investigate the function of RSG in
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Gibberellins (GAs) are important phytohormones for plant growth and development. DELLAs are members of the plant-specific GRAS protein family and act as repressors of GA signaling. DELLAs are rapidly degraded in the presence of GAs.... more
Gibberellins (GAs) are important phytohormones for plant growth and development. DELLAs are members of the plant-specific GRAS protein family and act as repressors of GA signaling. DELLAs are rapidly degraded in the presence of GAs. GA-GID1-DELLA complexes are recognized and ubiquitinated by the SCF(SLY) complex. The sleepy1 (sly1) F-box mutant exhibits dwarfism and low-germination phenotypes due to high accumulation of DELLAs. Overexpression of GID1 in the sly1 mutant partially rescues these phenotypes without degradation of DELLAs suggesting that proteolysis independent regulation of DELLAs exists in GA signaling. But the molecular mechanisms of non-proteolytic regulation of DELLA are largely unknown. Recently we identified a DELLA binding transcription factor, GAI-ASSOCIATED FACTOR1 (GAF1). GAF1 also interacts with co-repressor TOPLESS RELATED (TPR) in nuclei. DELLAs and TPR act as coactivator and corepressor of GAF1, respectively. GAs converts the GAF1 complex from transcription...
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A molecular mechanism to ensure signaling specificity is a scaffold. REPRESSION OF SHOOT GROWTH (RSG) is a tobacco (Nicotiana tabacum) transcription factor that is involved in gibberellin feedback regulation. The 14-3-3 proteins... more
A molecular mechanism to ensure signaling specificity is a scaffold. REPRESSION OF SHOOT GROWTH (RSG) is a tobacco (Nicotiana tabacum) transcription factor that is involved in gibberellin feedback regulation. The 14-3-3 proteins negatively regulate RSG by sequestering it in the cytoplasm in response to gibberellins. The N. tabacum Ca(2+)-dependent protein kinase NtCDPK1 was identified as an RSG kinase that promotes 14-3-3 binding of RSG by phosphorylation of RSG. CDPKs are unique sensor responders of Ca(2+) that are only found in plants and some protozoans. Here, we report a scaffolding function of CDPK. 14-3-3 proteins bound to NtCDPK1 by a new mode. Autophosphorylation of NtCDPK1 was necessary for the formation of the binding between NtCDPK1 and 14-3-3 but not for its maintenance. NtCDPK1 formed a heterotrimer with RSG and 14-3-3. Furthermore, we found that NtCDPK1 transfers 14-3-3 to RSG after phosphorylation of RSG and that RSG dissociates from NtCDPK1 as a complex with 14-3-3. ...
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Research Interests: Plant Biology, Tobacco, Regulation, Growth, Mutation, and 15 moreHistone Modification, Negative Feedback, Plant Growth Regulators, Feedback, Modification, Repression, Gibberellin, Transcription Factor, Homeostasis, Base Sequence, Histone, Gibberellins, PLANT PROTEINS, Shoot, and Molecular Sequence Data
Research Interests: Genetics, Plant Biology, Tobacco, Calcium, Protein Kinases, and 11 moreMutation, Arabidopsis, Substrate Specificity, Arginine, Calcium Binding Proteins, Amino Acid Substitution Rates, Structure activity Relationship, Protein Binding, PLANT PROTEINS, Biochemistry and cell biology, and Molecular Sequence Data
Research Interests: Genetics, Plant Biology, Tobacco, Transcription Factors, Humans, and 15 moreSequence alignment, Proteins, Plasmids, Arabidopsis, Plant cell, Amino Acid Sequence, Base Sequence, Recombinant Proteins, PLANT PROTEINS, Site-directed Mutagenesis, Protein Transport, Biochemistry and cell biology, Dimerization, Molecular Sequence Data, and conserved sequence
Research Interests: Genetics, Plant Biology, Tobacco, Proteins, Green Fluorescent Protein, and 11 moreEnzyme, Nicotiana Tabacum, Protein Degradation, Plant cell, Amino Acid Sequence, Protein Binding, Gibberellins, Transgenic plant, Biochemistry and cell biology, Molecular Sequence Data, and reverse transcriptase polymerase chain reaction
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Our study focused on the functional analysis of RSG, a tobacco transcriptional activator with a bZIP domain. Expression of a dominant-negative form of RSG severely inhibited the process of cell elongation in stems and reduced the... more
Our study focused on the functional analysis of RSG, a tobacco transcriptional activator with a bZIP domain. Expression of a dominant-negative form of RSG severely inhibited the process of cell elongation in stems and reduced the endogenous amounts of GA 1, which is the major active gibberellin (GA) in tobacco in transgenic plants. To investigate the function of RSG in
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Gibberellins (GAs) are essential regulators of plant development, and DELLAs are negative regulators of GA signaling. The mechanism of GA-dependent transcription has been explained by DELLA-mediated titration of transcriptional activators... more
Gibberellins (GAs) are essential regulators of plant development, and DELLAs are negative regulators of GA signaling. The mechanism of GA-dependent transcription has been explained by DELLA-mediated titration of transcriptional activators and their release through the degradation of DELLAs in response to GA. However, the effect of GA on genome-wide expression is predominantly repression, suggesting the existence of unknown mechanisms of GA function. In this study, we identified an Arabidopsis thaliana DELLA binding transcription factor, GAI-ASSOCIATED FACTOR1 (GAF1). GAF1 shows high homology to INDETERMINATE DOMAIN1 (IDD1)/ENHYDROUS. GA responsiveness was decreased in the double mutant gaf1 idd1, whereas it was enhanced in a GAF1 overexpressor. GAF1 binds to genes that are subject to GA feedback regulation. Furthermore, we found that GAF1 interacts with the corepressor TOPLESS RELATED (TPR) and that DELLAs and TPR act as coactivators and a corepressor of GAF1, respectively. GA conve...