Molecular studies on newly identified biomineralization genes in Paracentrotus lividus. (Abstract/Comunicazione in atti di convegno)

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Label
  • Molecular studies on newly identified biomineralization genes in Paracentrotus lividus. (Abstract/Comunicazione in atti di convegno) (literal)
Anno
  • 2011-01-01T00:00:00+01:00 (literal)
Alternative label
  • K. Karakostis, C. Costa, V. Matranga (2011)
    Molecular studies on newly identified biomineralization genes in Paracentrotus lividus.
    in Biomintec International Workshop, IBIM-CNR-Palermo, 7-8 febbraio 2011
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • K. Karakostis, C. Costa, V. Matranga (literal)
Pagina inizio
  • 24 (literal)
Pagina fine
  • 24 (literal)
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  • Biomintec International Workshop 2011 Palermo (literal)
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  • 1 (literal)
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  • Abstract (literal)
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  • IBIM - Istituto di Biomedicina e Immunologia Molecolare \"Alberto Monroy\" -Consiglio Nazionale delle Ricerche. Via Ugo La Malfa 153. - Palermo. (literal)
Titolo
  • Molecular studies on newly identified biomineralization genes in Paracentrotus lividus. (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#isbn
  • 9788890580505 (literal)
Abstract
  • The role of a set of genes in the formation of the magnesium calcite skeleton of Paracentrotus lividus (P. lividus) has been analyzed in this study. The group includes carbonic anhydrase (ca), tetraspanin (tetr), p16 (p16), p19 (p19) as well as the lectins: galectin (gal-8), advilin (adv), sm30a (sm30a) and sm50 (sm50). The coding sequences (CDS) were obtained by RT-PCR on the basis of in silico analysis and phylogenetic comparison with orthologous sequences. Comparative qPCR and whole mount in situ hybridization (WMISH) with Dig-labeled anti-sense DNA or RNA probes were employed for the temporal and spatial analysis of each transcript during development from fertilization to the late pluteus stage. Results demonstrated a modulation in the transcription levels of the genes which parallels skeleton development, increasing at the blastula stage and reaching multiple fold rises at the pluteus stage. In addition, we found that ca, adv, p16 and p19 were specifically expressed in one cellular population, the primary mesenchyme cells (PMCs), while tetr transcripts were localized within the ectoderm cell layer in the region facing the growing spicules and gal-8 is expressed in the ventral endoderm (foregut). Studies are in progress for the functional characterization of each protein coded by those genes. To this purpose, each CDS has been cloned in expression vectors for the preparation of recombinant proteins synthesized in vitro by E. coli BL21 (DE3). Protein expression and purification procedures have been successfully achieved for some of them. To study the effects of each protein on calcite formation, recombinant proteins will be used independently or in combination in biochemical assays, including in vitro crystallization experiments. In conclusion, a molecular tool-set of cDNAs and recombinant proteins is provided for further exploration of the calcification process in the sea urchin. References: - Zito F., C. Costa, S. Sciarrino, V. Poma, R. Russo, M. Angerer, V. Matranga (2003) Expression of univin, a TGF- growth factor, requires ectoderm-ECM interaction and promotes skeletal growth in the sea urchin embryo Dev Biol 264: 217-227 - Mann K., F.H. Wilt and A.J. Poustka, Mann et al. (2010) Proteomic analysis of sea urchin (Strongylocentrotus purpuratus) spicule matrix. Proteome Science 8: 33- - Wilt F. H. (1999) Matrix and Mineral in the Sea Urchin Larval Skeleton. J Struct Biol, 126:216-226 - Sea Urchin Genome Sequencing Consortium (2006). The genome of the sea urchin Strongylocentrotus purpuratus. Science 314: 941-952. - Illies M.R., Peeler MT, Dechtiaruk A.M., Ettensohn CA (2002) Identification and developmental expression of new biomineralization proteins in the sea urchin Strongylocentrotus purpuratus. - Dev. Genes Evol. 212:419-431 - Alvares K, Dixit SN, Lux E, and Veis A (2009) Echinoderm Phosphorylated Matrix Proteins UTMP16 and UTMP19 Have Different Functions in Sea Urchin. J Biol Chem 284: 26149-26160. - Zito F., Costa C., Sciarrino S., Cavalcante C.,Poma V., Matranga V. (2005) Cell Adhesion and Communication: A Lesson from Echinoderm Embryos for the Exploitation of New Therapeutic Tools, Progress in Molecular and Subcellular Biology Subseries Marine Molecular Biotechnology. V.Matranga (Ed.) Echinodermata Springer-Verlag Berlin Heidelberg (literal)
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