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

Mol Biol Rep. 2012 Mar;39(3):2633-44. doi: 10.1007/s11033-011-1016-7. Epub 2011 Jun 16.

Abstract

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 conserved 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' 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 putative regulatory regions located both in the upstream region and in the first intron containing putative binding sites for Forkhead and Nkx transcription factor families.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites / genetics
  • Computational Biology
  • DNA Footprinting
  • DNA Primers / genetics
  • Embryo, Nonmammalian*
  • Expressed Sequence Tags
  • Gene Expression Regulation / genetics*
  • Gene Transfer Techniques
  • Genes, Reporter / genetics
  • Genomics
  • Green Fluorescent Proteins / metabolism
  • Microinjections
  • Molecular Sequence Annotation
  • Nerve Tissue Proteins / genetics*
  • Paracentrotus / genetics*
  • Promoter Regions, Genetic / genetics*
  • Tubulin / genetics*

Substances

  • DNA Primers
  • Nerve Tissue Proteins
  • Tubulin
  • Green Fluorescent Proteins