Regulation of Arabidopsis thaliana Em genes: role of ABI5.
In order to identify new factors involved in Em (a class I Late Embryogenesis Abundant protein) gene expression, Arabidopsis mutants with an altered expression of an Em promoter GUS fusion construct and a modified accumulation of Em transcripts and proteins were isolated. Germination tests on ABA showed that the most affected mutant had a weak abi phenotype. Complementation tests further revealed this mutant to be a new abi5 allele, consequently named abi5-5. In addition to reducing the final level of Em transcripts in the dry seed, the abi5-5 mutation causes a delay in the accumulation of AtEm1 during seed development. An additional characteristic of the abi5-5 mutant, is the ability of its seeds to germinate at high concentrations of salt and mannitol. The abi5-5 mutation was characterized at the molecular level and was shown to result from a two base pair deletion in the coding sequence of the ABI 5 gene. The wild type and mutant recombinant proteins were produced in E. coli and were assayed for DNA-binding activity on their target promoters by electrophoretic mobility shift assay (EMSA). The ABI5 recombinant protein binds the ABRE sequence in the AtEm6 promoter as shown by Dnase footprinting. Among the ABRE-type sequences selected on both Em promoters, the G-box type AGACACGTGGCATGT element of the AtEm6 promoter shows the strongest binding by EMSA quantification.
Carles
Cristel
C
Laboratoire Génome et Développement des Plantes, Centre National de la Recherche Scientifique UMR 5096, Université de Perpignan, 52, Avenue de Villeneuve, 66860 Perpignan Cedex, France.
Bies-Etheve
Natacha
N
Aspart
Lorette
L
Léon-Kloosterziel
Karen M
KM
Koornneef
Maarten
M
Echeverria
Manuel
M
Delseny
Michel
M
D000040:Abscisic Acid / Q000494:pharmacology; D000483:Alleles; D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D024202:Electrophoretic Mobility Shift Assay; D018506:Gene Expression Regulation, Plant; D005816:Genetic Complementation Test; D018525:Germination / Q000187:drug effects; D016350:Leucine Zippers / Q000235:genetics; D008353:Mannitol / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010940:Plant Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012639:Seeds / Q000254:growth & development; D012965:Sodium Chloride / Q000494:pharmacology; D014157:Transcription Factors / Q000235:genetics* / Q000502:physiology
C409743:ABI5 protein, Arabidopsis; D029681:Arabidopsis Proteins; D050976:Basic-Leucine Zipper Transcription Factors; D004268:DNA-Binding Proteins; D010940:Plant Proteins; D014157:Transcription Factors; C066246:late embryogenesis abundant protein, plant; D008353:Mannitol; D012965:Sodium Chloride; D000040:Abscisic Acid