A G‐box motif (GCCACGTGCC) tetramer confers high‐level constitutive expression in dicot and monocot plants

Ishige, Fumiharu; Takaichi, Miyuki; Foster, Randy; Chua, Nam‐Hai; Oeda, Kenji

doi:10.1046/j.1365-313x.1999.00456.x

Cited by 99 publications

(85 citation statements)

References 33 publications

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“…Plant G-box elements contain a CACGTG core sequence that is necessary for efficient binding of basic leucine zipper proteins, and sequences flanking the CACGTG core affect basic leucine zipper protein binding specificity (25) and gene expression in vivo (26). The G-box in the cucumisin promoter, a 10-bp perfect palindrome (5Ј-GACACGTGTC-3Ј), can be classified as a high affinity class I G-box element that binds type A G-box binding factor (GBF) (16).…”

Section: Discussionmentioning

confidence: 99%

TGTCACA Motif Is a Novel cis-Regulatory Enhancer Element Involved in Fruit-specific Expression of the cucumisin Gene

Yamagata

Yonesu

Hirata

et al. 2002

Journal of Biological Chemistry

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Cucumisin, a subtilisin-like serine protease, is expressed at high levels in the fruit of melon (Cucumis melo L.) and accumulates in the juice. We investigated roles of the promoter regions and DNA-protein interactions in fruit-specific expression of the cucumisin gene. In transient expression analysis, a chimeric gene construct containing a 1.2-kb cucumisin promoter fused to a ␤-glucuronidase (GUS) reporter gene was expressed in fruit tissues at high levels, but the promoter activities in leaves and stems were very low. Deletion analysis indicated that a positive regulatory region is located between nucleotides ؊234 and ؊214 relative to the transcriptional initiation site. Gain-of-function experiments revealed that this 20-bp sequence conferred fruit specificity and contained a regulatory enhancer. Gel mobility shift experiments demonstrated the presence of fruit nuclear factors that interact with the cucumisin promoter. A typical G-box (GACACGTGTC) present in the 20-bp sequence did not bind fruit protein, but two possible cis-elements, an I-box-like sequence (AGATAT-GATAAAA) and an odd base palindromic TGTCACA motif, were identified in the promoter region between positions ؊254 and ؊215. The I-box-like sequence bound more tightly to fruit nuclear protein than the TGTCACA motif. The I-box-like sequence functions as a negative regulatory element, and the TGTCACA motif is a novel enhancer element necessary for fruit-specific expression of the cucumisin gene. Specific nucleotides responsible for the binding of fruit nuclear protein in these two elements were also determined.Timing and levels of gene expression are critical to the proper development of eukaryotic organisms. Regulation of the expression pattern of a particular gene can involve the specific binding of trans-acting factors to the cognate cis-elements, constituting a crucial step in transcriptional initiation and, in turn, on the spatial and temporal expression of genes. Plant genes that show tissue specificity, developmental specificity, and a wide range of expression levels have been characterized, whereas their expression patterns are also influenced by environmental stimuli. A family of genes for fruit proteins provide a model system for the study of the regulatory mechanisms of plant genes, since their expression is restricted to a specific tissue and stage during fruit development (1). A number of fruit-specific genes that are activated during ripening have been isolated from tomato and other fruits, and genes responding to ethylene and nonethylene signals have been identified (1, 2). The promoters of fruit-specific genes would also be of great interest for use in strategies to manipulate fruit metabolism and produce valuable proteins such as antibody, biopharmaceuticals, and edible vaccines through methods of genetic engineering (3-5). However, the detailed mechanisms by which the expression of fruit protein genes are regulated are poorly understood, as many of the essential cis-elements have not been identified.Melon cucumisin, an extracellular...

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Section: Discussionmentioning

confidence: 99%

TGTCACA Motif Is a Novel cis-Regulatory Enhancer Element Involved in Fruit-specific Expression of the cucumisin Gene

Yamagata

Yonesu

Hirata

et al. 2002

Journal of Biological Chemistry

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“…Experiments with a pG10-90:XVE..GUS transformant line, in which the ubiquitous G10-90 promoter (Ishige et al, 1999) drives inducible GUS expression, showed GUS expression that is highly dependent on 17-b-estradiol (Fig. 2, A and B).…”

Section: Stringency Of the Modified Inducible System In Transgenic Plmentioning

confidence: 99%

“…The XVE elements have been transferred into a single-site Gateway-compatible destination vector to allow highthroughput cloning of genes under the control of the inducible, ubiquitous promoter pG10-90 (Ishige et al, 1999;Curtis and Grossniklaus, 2003). Although useful, this vector does not allow easy cloning of other promoters to drive inducible expression.…”

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confidence: 99%

MultiSite Gateway-Compatible Cell Type-Specific Gene-Inducible System for Plants

et al. 2015

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A powerful method to study gene function is expression or overexpression in an inducible, cell type-specific system followed by observation of consequent phenotypic changes and visualization of linked reporters in the target tissue. Multiple inducible gene overexpression systems have been developed for plants, but very few of these combine plant selection markers, control of expression domains, access to multiple promoters and protein fusion reporters, chemical induction, and high-throughput cloning capabilities. Here, we introduce a MultiSite Gateway-compatible inducible system for Arabidopsis (Arabidopsis thaliana) plants that provides the capability to generate such constructs in a single cloning step. The system is based on the tightly controlled, estrogen-inducible XVE system. We demonstrate that the transformants generated with this system exhibit the expected cell type-specific expression, similar to what is observed with constitutively expressed native promoters. With this new system, cloning of inducible constructs is no longer limited to a few special cases but can be used as a standard approach when gene function is studied. In addition, we present a set of entry clones consisting of histochemical and fluorescent reporter variants designed for gene and promoter expression studies.

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“…The chimeric activator gene cassette contains the EcR ligand-binding region transcriptionally fused to the 441-bp GAL4 DNA-binding domain (amino acids 1-147; Laughon and Gesteland, 1984), the 264-bp VP16 acidic activation domain (amino acids 413-490) from Herpes simplex virus (Dalrymple et al, 1985), and the D to F regions containing the ligand-binding domain (amino acids 206-539) of EcR from Choristoneura fumiferana (spruce budworm; Perera et al, 1999), introduced between the constitutive G10-90 promoter (P 10-90 ; Ishige et al, 1999) and the nopaline synthase terminator sequence (Nos 3#). In p1002, the VP16 domain is positioned N terminally followed by the GAL4 and EcR domains (VGE), whereas in p1003, the GAL4 domain is positioned N terminally followed by the VP16 and EcR domains (GVE; Fig.…”

Section: Plasmid Constructsmentioning

confidence: 99%

Regulated Ethylene Insensitivity through the Inducible Expression of the Arabidopsis etr1-1 Mutant Ethylene Receptor in Tomato

Gallie

2010

Plant Physiol.

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Ethylene serves as an important hormone controlling several aspects of plant growth and development, including fruit ripening and leaf and petal senescence. Ethylene is perceived following its binding to membrane-localized receptors, resulting in their inactivation and the induction of ethylene responses. Five distinct types of receptors are expressed in Arabidopsis (Arabidopsis thaliana), and mutant receptors have been described that repress ethylene signaling in a dominant negative manner. One such mutant, ethylene resistant1-1 (etr1-1), results in a strong ethylene-insensitive phenotype in Arabidopsis. In this study, regulated expression of the Arabidopsis etr1-1 in tomato (Solanum lycopersicum) was achieved using an inducible promoter. In the absence of the inducer, transgenic seedlings remained sensitive to ethylene, but in its presence, a state of ethylene insensitivity was induced, resulting in the elongation of the hypocotyl and root in dark-grown seedlings in the presence of ethylene, a reduction or absence of an apical hook, and repression of ethylene-inducible E4 expression. The level of ethylene sensitivity could be controlled by the amount of inducer used, demonstrating a linear relationship between the degree of insensitivity and etr1-1 expression. Induction of etr1-1 expression also repressed the epinastic response to ethylene as well as delayed fruit ripening. Restoration of ethylene sensitivity was achieved following the cessation of the induction. These results demonstrate the ability to control ethylene responses temporally and in amount through the control of mutant receptor expression.

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A G‐box motif (GCCACGTGCC) tetramer confers high‐level constitutive expression in dicot and monocot plants

Cited by 99 publications

References 33 publications

TGTCACA Motif Is a Novel cis-Regulatory Enhancer Element Involved in Fruit-specific Expression of the cucumisin Gene

TGTCACA Motif Is a Novel cis-Regulatory Enhancer Element Involved in Fruit-specific Expression of the cucumisin Gene

MultiSite Gateway-Compatible Cell Type-Specific Gene-Inducible System for Plants

Regulated Ethylene Insensitivity through the Inducible Expression of the Arabidopsis etr1-1 Mutant Ethylene Receptor in Tomato

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