Hordein promoter methylation and transcriptional activity in wild-type and mutant barley endosperm
| Autor: | David J. Simpson, Mikael Blom Sørensen, Martin Müller, John H. Skerritt |
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| Rok vydání: | 1996 |
| Předmět: |
Glutens
Transcription Genetic Mutant Blotting Western Molecular Sequence Data GUS reporter system Biology Methylation Endosperm Epigenetics of physical exercise Gene Expression Regulation Plant Genes Reporter Genetics Cloning Molecular Promoter Regions Genetic Molecular Biology DNA Primers Glucuronidase Plant Proteins Regulation of gene expression Polymorphism Genetic Base Sequence food and beverages Promoter Hordeum Molecular biology Immunohistochemistry Blotting Southern DNA methylation Mutation |
| Zdroj: | Moleculargeneral genetics : MGG. 250(6) |
| ISSN: | 0026-8925 |
| Popis: | B- and C-hordein gene transcription is severely reduced in the endosperm of the regulatory barley mutant lys3a, and this is correlated with persistent hypermethylation of the promoters. In contrast, D-hordein is expressed at normal levels in the mutant. To confirm the connection between methylation and transcriptional activity, a genomic D-hordein clone was isolated and sequenced. The nucleotide composition of the promoter region revealed a CpG island and methylation analysis, using bisulphite treatment of genomic DNA, confirmed that the D-hordein promoter is unmethylated in endosperm and leaf tissue. Immunocytochemical studies localized D-hordein to the reticular component of protein bodies in both the wild-type Bomi and lys3a. Transient expression of GUS reporter gene constructs in barley endosperm, following transfection by particle bombardment revealed the D-hordein promotors. Comparison of transient expression in Bomi and lys3a endosperm demonstrated that the activities of the unmethylated D-hordein and the Hor1-14 C-hordein promoters were equivalent, while the activities in the mutant of the Horl-17 C-hordein and the Hor2-4 B-hordein promoters were reduced two- and tenfold, respectively. Methylation of plasmids in vitro prior to expression severely inhibited B- and D-hordein promoter activities. Based on these observations two categories of promoters for endosperm-specific expression of storage proteins are recognized and a model involving methylation and modulation of chromatin structure in the regulation by the Lys3 gene is presented. |
| Databáze: | OpenAIRE |
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