A 45‐bp proximal region containing AACA and GCN4 motif is sufficient to confer endosperm‐specific expression of the rice storage protein glutelin gene,<i>GluA‐3</i>

Yoshihara, Toshihiro; Washida, Haruhiko; Takaiwa, Fumio

doi:10.1016/0014-5793(96)00233-5

Cited by 52 publications

(27 citation statements)

References 37 publications

(51 reference statements)

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“…Actually, the new glutelin subunit GluB42 found in the course of genome projects is estimated to have a higher lysine content (4.21 mol%) than GluB2, which has the highest lysine content (3.60 mol%) among previously known subunits. However, the actual precise accumulation level of each gene product is so far difficult to predict by only gene sequences in spite of intensified promoter analysis [10][11][12][13], suggesting the importance and necessity of protein analysis. Besides, gene mining (cloning) by PCR based on DNA sequence homology sometimes encounters a difficulty [14] and alternative methods at protein level such as screening with expression library and antibody may be rather useful [15], especially when analyzed on distant relative species.…”

Section: Introductionmentioning

confidence: 98%

Diversity of rice glutelin polypeptides in wild species assessed by the higher‐temperature sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and subunit‐specific antibodies

et al. 2008

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In efforts to find genetic resources with high nutritional value of rice seed, we assessed the diversity of the major storage protein glutelin in 13 wild and 2 cultivated rice species by a unique SDS-PAGE method and subunit-specific antibodies. Maximum separation of microheterogeneous glutelin alpha-polypeptides, which is a prerequisite for the diversity evaluation, could be attained by SDS-PAGE performed at higher temperature (45 degrees C) than the generally employed temperatures (4-25 degrees C). Seven antipeptide antibodies were raised against subunit-specific epitope sequences designed at five sites from four variable regions spanning the glutelin alpha-polypeptides. High specificity of each antibody was confirmed using rice glutelin mutants, and demonstrated considerable variation in amino acid sequence and accumulation level of glutelin subunit in wild species, in combination with the higher-temperature SDS-PAGE. The degree of the variation was, however, changed according to the site of variable regions and the type of subunit. Some wild species accumulated nutritious GluB subunits more than cultivated rice. The wild species Oryza longiglumis and O. brachyantha had glutelin with low reactivity against most antibodies examined in this study, reflecting the significant divergence. Such wild species may hopefully serve as important genetic resources for nutritional improvement of cultivated rice.

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Section: Introductionmentioning

confidence: 98%

Diversity of rice glutelin polypeptides in wild species assessed by the higher‐temperature sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and subunit‐specific antibodies

et al. 2008

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“…In most rice glutelin genes, these two motifs are also associated with the AACA motif (12). Gain-of-function experiments showed that combinations of two motifs (GCN4 motif and prolamin box, GCN4 motif and AACA motif) are not sufficient to confer endosperm-specific expression, suggesting that participation of additional element is required to form a functional complex of trans-acting factors (6,13). It has been recently demonstrated in the rice glutelin GluB-1 gene that at least three cis-elements containing the GCN4, ACGT, and AACA motifs within the * This work was supported by research grants from the Bio-oriented Technology Research Advancement Institute (PROBRAIN), Science and Technology Agency (Enhancement System of Center of Excellence), and the Ministry of Agriculture, Forestry and Fishery of Japan (to F. T.).…”

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A Rice Functional Transcriptional Activator, RISBZ1, Responsible for Endosperm-specific Expression of Storage Protein Genes through GCN4 Motif

Onodera¹,

Suzuki²,

Wu³

et al. 2001

Journal of Biological Chemistry

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149

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The GCN4 motif, a cis-element that is highly conserved in the promoters of cereal seed storage protein genes, plays a central role in controlling endospermspecific expression. This motif is the recognition site for a basic leucine zipper transcriptional factor that belongs to the group of maize Opaque-2 (O2)-like proteins. Five different basic leucine zipper cDNA clones, designated RISBZ1-5, have been isolated from a rice seed cDNA library. The predicted gene products can be divided into two groups based on their amino acid sequences. Although all the RISBZ proteins are able to interact with the GCN4 motif, only RISBZ1 is capable of activating (more than 100-fold expression) the expression of a reporter gene under a minimal promoter fused to a pentamer of the GCN4 motif. Loss-of-function and gain-of-function experiments using the yeast GAL4 DNA binding domain revealed that the proline-rich N-terminal domain (27 amino acids in length) is responsible for transactivation. The RISBZ1 protein is capable of forming homodimers as well as heterodimers with other RISBZ subunit proteins. RISBZ1 gene expression is restricted to the seed, where it precedes the expression of storage protein genes. When the RISBZ1 promoter was transcriptionally fused to the ␤-glucuronidase reporter gene and the chimeric gene was introduced into rice, the ␤-glucuronidase gene is specifically expressed in aleurone and subaleurone layer of the developing endosperm. These findings suggest that the specific expression of transcriptional activator RISBZ1 gene may determine the endosperm specificity of the storage protein genes.Regulated gene expression is mediated by the combinatorial interactions of multiple cis-elements in the gene's promoter. Specific binding of transcriptional factors to the cognate ciselements constitute a crucial step in transcription initiation and, in turn, on the spatial and temporal expression of genes.Seed storage protein genes provide a model system for the study on the regulatory mechanisms of plant genes (1), since their expression is restricted to a specific tissue and stage during seed development. These specific temporal and spatial expression patterns may be explained as the result of regulatory assemblies of several transcriptional activators that recognize the cis-elements implicated in seed-specific expression. Therefore, to understand such molecular mechanisms, characterization of cis-elements and transcription factors has been performed on many storage protein genes of several crop plants (2,3). Despite numerous studies, the mechanism by which these genes are regulated are poorly understood, since many of the essential cis-elements have not been identified. This is especially true in the case of monocot plants, where many of the promoter analyses of cereal storage protein genes have carried out by transient assays using particle bombardment or heterologous transgenic tobacco system (4 -6). Dissection analyses of promoter using homologous stable transgenic plant have been carried out only on glutelin genes of ...

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“…These are the GCN4-like motif (GLM) and the prolamin box (PB), which together constitute the endosperm box (EB; Colot et al, 1989), and the AACA motif (Zheng et al, 1993). The PB and the AACA motif confer endosperm-specific expression (Colot et al, 1987;Yoshihara et al, 1996), whereas GLM is thought to play a role in the nutritional regulation of prolamin gene expression (Mü ller and Knudsen, 1993).…”

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Nucleotide Polymorphism in the Wheat Transcriptional ActivatorSpaInfluences Its Pattern of Expression and Has Pleiotropic Effects on Grain Protein Composition, Dough Viscoelasticity, and Grain Hardness

et al. 2009

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Storage protein activator (SPA) is a key regulator of the transcription of wheat (Triticum aestivum) grain storage protein genes and belongs to the Opaque2 transcription factor subfamily. We analyzed the sequence polymorphism of the three homoeologous Spa genes in hexaploid wheat. The level of polymorphism in these genes was high particularly in the promoter. The deduced protein sequences of each homoeolog and haplotype show greater than 93% identity. Two major haplotypes were studied for each Spa gene. The three Spa homoeologs have similar patterns of expression during grain development, with a peak in expression around 300 degree days after anthesis. On average, Spa-B is 10 and seven times more strongly expressed than Spa-A and Spa-D, respectively. The haplotypes are associated with significant quantitative differences in Spa expression, especially for Spa-A and Spa-D. Significant differences were found in the quantity of total grain nitrogen allocated to the gliadin protein fractions for the Spa-A haplotypes, whereas the synthesis of glutenins is not modified. Genetic association analysis between Spa and dough viscoelasticity revealed that Spa polymorphisms are associated with dough tenacity, extensibility, and strength. Except for Spa-A, these associations can be explained by differences in grain hardness. No association was found between Spa markers and the average single grain dry mass or grain protein concentration. These results demonstrate that in planta Spa is involved in the regulation of grain storage protein synthesis. The associations between Spa and dough viscoelasticity and grain hardness strongly suggest that Spa has complex pleiotropic functions during grain development.

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A 45‐bp proximal region containing AACA and GCN4 motif is sufficient to confer endosperm‐specific expression of the rice storage protein glutelin gene,GluA‐3

Cited by 52 publications

References 37 publications

Diversity of rice glutelin polypeptides in wild species assessed by the higher‐temperature sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and subunit‐specific antibodies

Diversity of rice glutelin polypeptides in wild species assessed by the higher‐temperature sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and subunit‐specific antibodies

A Rice Functional Transcriptional Activator, RISBZ1, Responsible for Endosperm-specific Expression of Storage Protein Genes through GCN4 Motif

Nucleotide Polymorphism in the Wheat Transcriptional ActivatorSpaInfluences Its Pattern of Expression and Has Pleiotropic Effects on Grain Protein Composition, Dough Viscoelasticity, and Grain Hardness

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