6533b861fe1ef96bd12c45d3

RESEARCH PRODUCT

In silico characterization of the neural alpha tubulin gene promoter of the sea urchin embryo Paracentrotus lividus by phylogenetic footprinting

Maria Antonietta RagusaMarco EmanueleSalvatore CostaFabrizio GianguzzaValeria Longo

subject

Embryo NonmammalianMicroinjectionsSequence analysisGreen Fluorescent ProteinsDNA FootprintingNerve Tissue ProteinsSettore BIO/11 - Biologia MolecolarePhylogenetic footprintingParacentrotus lividusGenes ReporterTubulinGeneticsAnimalsPromoter Regions GeneticMolecular BiologyGeneDNA PrimersExpressed Sequence TagsComparative genomicsGeneticsBinding SitesbiologyGene Transfer TechniquesComputational BiologyMolecular Sequence AnnotationPromoterGenomicsGeneral MedicineSea urchin Neural development Gene expression Phylogenetic footprint Cis-regulatory analysisbiology.organism_classificationGene Expression RegulationRegulatory sequenceParacentrotusOrthologous Gene

description

During Paracentrotus lividus sea urchin embryo development one alpha and one beta tubulin genes are expressed specifically in the neural cells and they are early end output of the gene regulatory network that specifies the neural commitment. In this paper we have used a comparative genomics approach to identify con- served regulatory elements in the P. lividus neural alpha tubulin gene. To this purpose, we have first isolated a genomic clone containing the entire gene plus 4.5 Kb of 5 0 upstream sequences. Then, we have shown by gene transfer experiments that its non-coding region drives the spatio- temporal gene expression corresponding substantially to that of the endogenous gene. In addition, we have identified by genome and EST sequence analysis the S. purpuratus alpha tubulin orthologous gene and we propose a revised annotation of some tubulin family members. Moreover, by computational techniques we delineate at least three puta- tive regulatory regions located both in the upstream region and in the first intron containing putative binding sites for Forkhead and Nkx transcription factor families.

https://doi.org/10.1007/s11033-011-1016-7