Characterization and Promoter Analysis of a Cotton RING-Type Ubiquitin Ligase (E3) Gene
| Autor: | Johnie N. Jenkins, Sukumar Saha, Meng-Hsuan Ho, Din-Pow Ma |
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| Rok vydání: | 2010 |
| Předmět: |
Ubiquitin-Protein Ligases
Arabidopsis Bioengineering GUS reporter system Genes Plant Applied Microbiology and Biotechnology Biochemistry Gene Expression Regulation Plant Complementary DNA Gene expression Ring finger medicine Amino Acid Sequence Cotton Fiber Cloning Molecular Promoter Regions Genetic Molecular Biology Zinc finger Regulation of gene expression Gossypium Sequence Homology Amino Acid biology Ubiquitination Sequence Analysis DNA Plants Genetically Modified Molecular biology Protein ubiquitination Ubiquitin ligase medicine.anatomical_structure biology.protein Biotechnology |
| Zdroj: | Molecular Biotechnology. 46:140-148 |
| ISSN: | 1559-0305 1073-6085 |
| DOI: | 10.1007/s12033-010-9280-7 |
| Popis: | A cotton fiber cDNA, GhRING1, and its corresponding gene have been cloned and characterized. The GhRING1 gene encodes a RING-type ubiquitin ligase (E3) containing 338 amino acids (aa). The GhRING1 protein contains a RING finger motif with conserved cysteine and histine residues at the C-terminus, and is classified as a C(3)H(2)C(3)-type RING protein. Blast searches show that GhRING1 has the highest homology to At3g19950, a zinc finger family protein from Arabidopsis. Real time RT-PCR analysis indicates that the GhRING1 gene is expressed in cotton fibers in a developmental manner. The transcript level of GhRING1 gene reaches a maximum in elongating fibers at 15 days post-anthesis (DPA). In vitro auto-ubiquitination assays using wheat germ extract and a reconstitution system demonstrate that GhRING1 has the ubiquitin E3 ligase activity. The histochemical GUS assay was performed to analyze tissue specificity of the GhRING1 and At3g19950 promoters in transgenic Arabidopsis plants. The GUS assay shows that the promoter of At3g19950 is highly activated in leaves, roots, trichomes, and also in anthers and stigma of flowers. In contrast, the GUS expression directed by the GhRING1 promoter is only located at stipules and anthers. The expression pattern of GhRING1 suggests that protein ubiquitination and turnover may be involved in transition to different stages of cotton fiber development. |
| Databáze: | OpenAIRE |
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