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X W Deng

Publications and source records attributed to X W Deng.

At least 55 records · Page 3Linked to original sources
PubMed1998

Combinatorial interaction of light-responsive elements plays a critical role in determining the response characteristics of light-regulated promoters in Arabidopsis.

We have studied the roles of PhyA, PhyB and CRY1 photoreceptors and the downstream light-signaling components, COP1 and DET1, in mediating high-irradiance light-controlled activity of promoters containing synthetic light-responsive elements (LRE). Promoters with paired LREs were able to respond to a wide spectrum of light through multiple photoreceptors, while the light-inducible single LRE promoters primarily responded to a specific wavelength of light. In addition, our results indicate that Cry1 is involved in PhyB-mediated red-light induction of the G-GATA/NOS101 promoter, and that both Cry1 and PhyB are required for effective repression of the GT1/NOS101 promoter by red or blue light. An interaction between PhyA and PhyB in mediating GT1-GATA/NOS101 promoter light activation was also observed. Furthermore, our data indicate that COP1 and DET1 exert negative control in the dark only on paired LRE promoters but not single LRE promoters. From these results, we conclude that the combinatorial interaction of LREs is essential in determining the ability of light-responsive promoters to be modulated by crucial cellular regulators and to respond to diverse light environments.

Arabidopsis↗
PubMed1998

Multiple photoreceptors mediate the light-induced reduction of GUS-COP1 from Arabidopsis hypocotyl nuclei.

The subcellular localization of COP1, a key photomorphogenic repressor, is regulated by light in Arabidopsis seedlings. Photoreceptor loss-of-function mutants and dominant gain-of-function overexpression transgenes were both used to analyze the influences of the three photoreceptors, phyA, phyB, and CRY1, on the light-regulated subcellular localization of COP1. Through a semiquantitative analysis of the nuclear abundance of GUS-COP1 in the various genetic backgrounds, the specific roles of the individual photoreceptors have been established. The data suggest that multiple photoreceptors influence the light-regulated subcellular localization of COP1 in white light. Under specific wavelengths of light, phyA, phyB, and CRY1 each play critical roles in mediating far-red, red, and blue light signals, respectively. Our data also support an interdependency between CRY1 and the phytochromes in mediating the light-regulated subcellular localization of COP1 and thus seedling development.

Arabidopsis↗
PubMed1998

Arabidopsis homologs of a c-Jun coactivator are present both in monomeric form and in the COP9 complex, and their abundance is differentially affected by the pleiotropic cop/det/fus mutations.

The CONSTITUTIVE PHOTOMORPHOGENIC9 (COP9) complex is a nuclear localized, multisubunit protein complex essential for repression of light-mediated development in Arabidopsis. Mutations that abolish the complex result in constitutive photomorphogenic development in darkness and pleiotropic developmental defects in both light and darkness. Here, we report the identification of two apparently redundant genes, AJH1 and AJH2, that encode a subunit of the COP9 complex. Both AJH1 and AJH2 share high amino acid sequence identity (62 and 63%, respectively) with JAB1, a specific mammalian coactivator of AP-1 transcription. The proteins encoded by these two genes are present in both complex and monomeric forms, whereas complex formation is in part mediated by the direct interaction with FUSCA6. In addition, the stability of the monomeric AJH proteins requires functional COP1 and DEETIOLATED1 loci. Together with the fact that the previously known subunit FUSCA6 is an Arabidopsis homolog of human GPS1, a negative regulator of AP-1 transcription, our data suggest that the COP9 complex may contain both negative and positive regulators of transcription. Therefore, the COP9 complex may achieve its pleiotropic effects on Arabidopsis development by modulating activities of transcription factors in response to environmental stimuli.

Amino Acid Sequence↗
PubMed1998

Arabidopsis bZIP protein HY5 directly interacts with light-responsive promoters in mediating light control of gene expression.

The Arabidopsis HY5 gene has been defined genetically as a positive regulator of photomorphogenesis and recently has been shown to encode a basic leucine zipper type of transcription factor. Here, we report that HY5 is constitutively nuclear localized and is involved in light regulation of transcriptional activity of the promoters containing the G-box, a well-characterized light-responsive element (LRE). In vitro DNA binding studies suggested that HY5 can bind specifically to the G-box DNA sequences but not to any of the other LREs present in the light-responsive promoters examined. High-irradiance light activation of two synthetic promoters containing either the consensus G-box alone or the G-box combined with the GATA motif (another LRE) and the native Arabidopsis ribulose bisphosphate carboxylase small subunit gene RBCS-1A promoter, which has an essential copy of the G-box, was significantly compromised in the hy5 mutant. The hy5 mutation's effect on the high-irradiance light activation of gene expression was observed in both photosynthetic and nonphotosynthetic tissues. Furthermore, the characteristic phytochrome-mediated red light- and far-red light-reversible low-fluence induction of the G-box-containing promoters was diminished specifically in hy5 plants. These results suggest that HY5 may interact directly with the G-box in the promoters of light-inducible genes to mediate light-controlled transcriptional activity.

Arabidopsis↗
PubMed1998

Role of a COP1 interactive protein in mediating light-regulated gene expression in arabidopsis.

Arabidopsis seedlings display distinct patterns of gene expression and morphogenesis according to the ambient light condition. An Arabidopsis nuclear protein, CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1), acts to repress photomorphogenesis in the absence of light. The Arabidopsis CIP7 protein was identified by its capability to interact with COP1. CIP7 is a novel nuclear protein that contains transcriptional activation activity without a recognizable DNA binding motif. CIP7 requires light for its high level of expression, and COP1 seems to play a role in repressing its expression in darkness. Decreasing CIP7 expression by introducing antisense CIP7 RNA resulted in defects in light-dependent anthocyanin and chlorophyll accumulation. Antisense plants also displayed reduced expression of light-inducible genes for anthocyanin biosynthesis and photosynthesis. However, no defect was observed in light-dependent inhibition of hypocotyl elongation. Taken together, our data indicate that CIP7 acts as a positive regulator of light-regulated genes and is a potential direct downstream target of COP1 for mediating light control of gene expression.

Amino Acid Sequence↗
PubMed1998

Characterization and purification of the mammalian COP9 complex, a conserved nuclear regulator initially identified as a repressor of photomorphogenesis in higher plants.

The COP9 complex has been identified as a repressor of photomorphogenesis in Arabidopsis. Here we demonstrate that the COP9 complex is also present in mammals. Specific antibodies were generated against human counterparts of the Arabidopsis COP9 and COP11, the two known subunits of plant COP9 complex. Using these antibodies, we showed that indeed mammalian COP9 and COP11, also known as GPS1, could be coimmuno-precipitated using either of the two specific antibodies, definitively confirming that they are physically part of the same complex. Further, the mammalian COP9 and COP11/GPS1 were cofractionated in the same large molecular weight fractions of about 500 kDa and were absent from the monomeric fractions. The mammalian COP9 complex was present in all organs examined but abundances vary. Indirect immunofluorescence studies suggested that the mammalian COP9 complex is largely nuclear localized. Both conventional biochemical and affinity purifications of the COP9 complex from pig spleen indicated that the mammalian COP9 complex consists of eight distinct subunits. These findings indicate that mammals also have a COP9 complex with conserved molecular composition and biochemical and cellular properties similar to the higher plant counterpart.

Animals↗
PubMed1998

Light control of Arabidopsis developmental pattern.

As sessile organisms, higher plants are characterized by a high degree of developmental pattern plasticity in response to environmental signals, and in many cases respond to the changing environment by tailoring their developmental patterns in a way that maximizes their chances of survival and reproduction. Given the importance of photosynthesis to plant survival, light signals are arguably among the most important environmental cues to plant development. Genetic analysis of the light control of the Arabidopsis seedling development pattern has revealed that the pleiotropic COP/DET/FUS genes play a key role in integrating light signals and modulating developmental pattern formation. Recent studies support a working model in which COP1 act within the nucleus to sequester and inactivate transcription factors in darkness, while light abrogates this association by modulating COP1 nuclear abundance. This results in activation of the transcription factors and expression of genes responsible for photomorphogenic development. The other pleiotropic COP/DET/FUS proteins act to maintain the proper nuclear localization or retention of COP1 in darkness.

Arabidopsis↗
PubMed1997

Genetic and developmental control of nuclear accumulation of COP1, a repressor of photomorphogenesis in Arabidopsis.

Using a beta-glucuronidase (GUS) reporter-COP1 fusion transgene, it was shown previously that Arabidopsis COP1 acts within the nucleus as a repressor of seedling photomorphogenic development and that high inactivation of COP1 was accompanied by a reduction of COP1 nuclear abundance (A.G. von Arnim, X.-W. Deng [1994] Cell 79: 1035-1045). Here we report that the GUS-COP1 fusion transgene can completely rescue the defect of cop1 mutations and thus is fully functional during seedling development. The kinetics of GUS-COP1 relocalization in a cop1 null mutant background during dark/light transitions imply that the regulation of the functional nuclear COP1 level plays a role in stably maintaining a committed seedling's developmental fate rather than in causing such a commitment. Analysis of GUS-COP1 cellular localization in mutant hypocotyls of all pleiotropic COP/DET/FUS loci revealed that nuclear localization of GUS-COP1 was diminished under both dark and light conditions in all mutants tested, whereas nuclear localization was not affected in the less pleiotropic cop4 mutant. Using both the brassinosteroid-deficient mutant det2 and brassinosteroid treatment of wild-type seedlings, we have demonstrated that brassinosteroid does not control the hypocotyl cell elongation through regulation nuclear localization of COP1. The growth regulator cytokinin, which also dramatically reduced hypocotyl cell elongation in the absence of light, did not prevent GUS-COP1 nuclear localization in dark-grown seedlings. Our results suggest that all of the previously characterized pleiotropic COP/DET/FUS loci are required for the proper nuclear localization of the COP1 protein in the dark, whereas the less pleiotropic COP/DET loci or plant regulators tested are likely to act either downstream of COP1 or by independent pathways.

Arabidopsis↗
PubMed1996

Combinatorial interplay of promoter elements constitutes the minimal determinants for light and developmental control of gene expression in Arabidopsis.

Higher plants are able to integrate environmental and endogenous signals to regulate gene expression for optimal development. To define the minimal sequence requirement sufficient to integrate light and developmental signals in controlling promoter activity, we carried out a systematic analysis of the roles of four well-conserved 'light-responsive elements (LREs)' common to many nuclear-encoded photosynthetic genes. A gain-of-function assay using basal promoter-reporter fusions in stable transgenic Arabidopsis was employed to demonstrate that pairwise combinations of the LREs, but not the individual elements alone, can confer light-inducible expression to the reporter gene independently of the basal promoter context and the light-triggered morphological changes. The activity of the synthetic promoters with the paired LREs can be modulated at least by the phytochrome system. Further, those synthetic light-regulated promoters confer a photosynthetic cell-specific expression pattern and respond to the chloroplast development state. Our data suggest that distinct combinatorial interactions of LREs can serve as minimal autonomous promoter determinants which integrate light and developmental signals and modulate promoter activity.

Arabidopsis↗
PubMed1996

The COP9 complex, a novel multisubunit nuclear regulator involved in light control of a plant developmental switch.

Arabidopsis COP9 is a component of a large protein complex that is essential for the light control of a developmental switch and whose conformation or size is modulated by light. The complex is acidic, binds heparin, and is localized within the nucleus. Biochemical purification of the complex to near homogeneity revealed that it contains 12 distinct subunits. One of the other subunits is COP11, mutations in which result in a phenotype identical to cop9 mutants. The COP9 complex may act to regulate the nuclear abundance of COP1, an established repressor of photomorphogenic development. During the biogenesis of the COP9 complex, a certain degree of prior subunit association is a prerequisite for proper nuclear translocation. Since both COP9 and COP11 have closely related human counterparts, the COP9 complex probably represents a conserved developmental regulator in higher eukaryotes.

Arabidopsis↗
PubMed1996

A role for transcriptional repression during light control of plant development.

Light mediates plant development partly by orchestrating changes in gene expression, a process which involves a complex combination of positive and negative signaling cascades. Genetic investigations using the small crucifer Arabidopsis thaliana have demonstrated a fundamental role for the down-regulation of light-inducible genes in response to darkness, thus offering a suitable model system for investigating how plants repress gene expression in a developmental context. Rapid progress in eukaryotic gene repression mechanisms in general, and light control of plant gene expression in particular, sheds new light on how a class of ten pleiotropic COP/DET/FUS genes might function to down-regulate light-inducible genes in plants.

Arabidopsis↗
PubMed1996

A complement of ten essential and pleiotropic arabidopsis COP/DET/FUS genes is necessary for repression of photomorphogenesis in darkness.

Two genetic screens, one for mutations resulting in photomorphogenic development in darkness and the other for mutants with fusca phenotype, have thus far identified six pleiotropic Arabidopsis COP/DET/FUS genes. Here, we characterized representative mutants that define four additional pleiotropic photomorphogenic loci and a null mutant allele of the previously defined DET1 locus. Dark-grown seedlings homozygous for these recessive mutations exhibit short hypocotyls and expanded cotyledons and are lethal before reaching reproductive development. Dark-grown mutant seedlings also display characteristic photomorphogenic cellular differentiation and elevated expression of light-inducible genes. In addition, analyses of plastids from dark-grown mutants reveal partial chloroplast differentiation and absence of etioplast development. Root vascular bundle cells of light-grown mutant seedlings develop chloroplasts, suggesting that these FUS gene products are important for suppression of chloroplast differentiation in light-grown roots. Double-mutant analyses indicate that these pleiotropic cop/det/fus mutations are epistatic to mutations in phytochromes, a blue-light photoreceptor, and a downstream regulatory component, HY5. Therefore, there is a complement of at least 10 essential and pleiotropic Arabidopsis genes that are necessary for repression of photomorphogenic development.

Arabidopsis↗
PubMed1996

Evidence for FUS6 as a component of the nuclear-localized COP9 complex in Arabidopsis.

The pleiotropic CONSTITUTIVE PHOTOMORPHOGENIC (COP), DEETIOLATED (DET), and FUSCA (FUS) loci are essential regulatory genes involved in the light control of seedling developmental patterns in Arabidopsis. Although COP1, DET1, COP9, and FUS6 (also called COP11) have been cloned, their biochemical activities and interactions remain elusive. We have recently suggested that multiple pleiotropic COP, DET, and FUS genes may encode subunits of a large regulatory complex. In this study, we generated specific antibodies against Arabidopsis FUS6 and show that accumulation of both COP9 and FUS6 is coordinated in the pleiotropic cop, det, and fus mutant backgrounds and in wild-type plants throughout development. Both COP9 and FUS6 cofractionated into identical high molecular mass fractions in an analytical gel filtration assay, and neither was found in its monomeric form. Moreover, antibodies raised against either COP9 or FUS6 selectively coimmunoprecipitated both proteins. We have also developed an Arabidopsis protoplast immunolocalization assay and demonstrated that the COP9 complex is localized in the nucleus and that its nuclear localization is not affected by light conditions or tissue types. The integrated genetic and biochemical results strongly support the conclusion that both COP9 and FUS6 are components of the nuclear-localized COP9 complex. Therefore, we have provided the strongest evidence for the conclusion that at least some of the pleiotropic COP, DET, and FUS loci act in the same signaling pathway.

Arabidopsis↗
PubMed1996

Expression of an N-terminal fragment of COP1 confers a dominant-negative effect on light-regulated seedling development in Arabidopsis.

CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1) is an essential regulatory gene that plays a role in light control of seedling development in Arabidopsis. The COP1 protein possesses three recognizable structural domains: a RING finger zinc binding domain near the N terminus, followed by a coiled-coll domain and a domain with WD-40 repeats in the C-terminal half. To determine whether COP1 acts specifically as a light-inactivable repressor of photomorphogenic development and to elucidate the functional roles of the specific structural domains, mutant cDNAs encoding the N-terminal 282 amino acids (N282) of COP1 were expressed and analyzed in transgenic plants. High-level expression of the N282 fragment caused a dominant-negative phenotype similar to that of the loss-of-function cop1 mutants. The phenotypic characteristics include hypersensitivity of hypocotyl elongation to inhibition by white, blue, red, and far-red light stimuli. In the dark, N282 expression led to pleiotropic photomorphogenic cotyledon development, including cellular differentiation, plastid development, and gene expression, although it has no significant effect on the hypocotyl elongation. However, N282 expression had a minimal effect on the expression of stress- and pathogen-inducible genes. These observations support the hypothesis that COP1 is directly involved in the light control of seedling development and that it acts as a repressor of photomorphogenesis. Further, the results imply that the N282 COP1 fragment, which contains the zinc binding and colled-coil domains, is capable of interacting with either downstream targets or with the endogenous wild-type COP1, thus interfering with normal regulatory processes. The fact the N282 is able to interact with N282 and full-length COP1 in yeast provided evidence for the latter possibility.

Arabidopsis↗
PubMed1995

Arabidopsis COP1 protein specifically interacts in vitro with a cytoskeleton-associated protein, CIP1.

Arabidopsis COP1 acts inside the nucleus to suppress photomorphogenic cellular development, and light inactivation of COP1 may involve a specific control of its nuclear activity in hypocotyls and cotyledons, but not in roots, of developing seedlings. To understand the molecular mechanisms of COP1 action during light-mediated development, we initiated a screen for Arabidopsis cDNAs encoding proteins which interact directly with COP1 in vitro as a step to identify the cellular components involved. We report here the isolation and characterization of a cDNA clone encoding a protein designated CIP1 (COP1-interactive protein 1). CIP1 is predominantly alpha-helical and most likely involved in coiled-coil formation. It interacts specifically with the putative coiled-coil region of COP1 in vitro. Further, CIP1 is encoded by a single gene in Arabidopsis, and its mRNA and protein levels are not regulated by light. Immunofluorescent labeling of CIP1 in Arabidopsis seedling protoplasts demonstrated that CIP1 is part of, or associated with, a cytoskeletal structure in hypocotyl and cotyledon cells, but not in roots. Our results are consistent with a possible role of CIP1 in mediating light control of COP1 nuclear activity by regulating its nucleocytoplasmic partitioning.

Amino Acid Sequence↗