ABSTRACT
UNLABELLED: Grass seeds are complex organs composed by multiple tissues and cell types that develop coordinately to produce a viable embryo. The identification of genes involved in seed development is of great interest, but systematic spatial analyses of gene expression on maize seeds at the cell level have not yet been performed. MASISH is an online database holding information for gene expression spatial patterns in maize seeds based on in situ hybridization experiments. The web-based query interface allows the execution of gene queries and provides hybridization images, published references and information of the analyzed genes. AVAILABILITY: http://masish.uab.cat/.
Subject(s)
Computational Biology/methods , Databases, Genetic , Gene Expression Regulation, Plant , Seeds/genetics , Zea mays/genetics , Gene Expression Profiling , Internet , SoftwareABSTRACT
The maize genomic DNA sequence encoding the eukaryotic translation initiation factor 5 (eIF-5) has been isolated from genomic library of maize seedlings and the exon-intron structure determined (accession number AJ132240). The length of genomic DNA sequenced was about 7kb and contained two exons with the translation start site in exon 2. The only intron is located in the non-coding 5' region and it is 1298bp long with the splice acceptor and donor sites conforming to the AG/GT rules. Repetitive sequence fragments are located in the 5' and 3' intergenic region. The accumulation of eIF-5 mRNA was studied by RNA blot and in situ hybridization. The observed distribution of mRNA may correlate with the function of the protein, as it appears to be highly abundant in tissues where the proportion of cells actively dividing is very high, such as meristematic regions.
Subject(s)
Genes, Plant/genetics , Peptide Initiation Factors/genetics , Zea mays/genetics , Cloning, Molecular , DNA, Plant/chemistry , DNA, Plant/genetics , Eukaryotic Initiation Factor-5 , Exons , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , In Situ Hybridization , Introns , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , Seeds/genetics , Sequence Analysis, DNA , Tissue Distribution , Zea mays/chemistry , Zea mays/embryologyABSTRACT
A maize cDNA encoding the eukaryotic translation initiation factor 5 (eIF-5) has been isolated from an 8-day-old seedling cDNA library. The 1975 bp cDNA encodes a protein of 451 amino acids, with a predicted molecular weight of 49.04 kDa, and hybridizes to a single sequence in the maize genome. The deduced sequence contains motifs characteristic of proteins belonging to the GPTase superfamily, a zinc finger well conserved in all the protein sequences for eIF-5 reported so far, and a fragment also present in prokaryotic and chloroplast L11 ribosomal protein. Polymer-binding assays have been used to assess the predicted RNA binding property of the protein and to characterize its function. It is shown that the eIF-5-encoded protein binds to single-stranded DNA and to polyuridylic acid and that the binding is dependent on the presence of Zn2+ ions. These results suggest that the zinc-finger structure is involved in the binding of the eIF-5 protein to RNA.