Transcriptional increase and misexpression of 14-3-3 epsilon in sea urchin embryos exposed to UV-B.
Members of the 14-3-3 protein family are involved in many important cellular events, including stress response, survival and apoptosis. Genes of the 14-3-3 family are conserved from plants to humans, and some members are responsive to UV radiation. Here, we report the isolation of the complete cDNA encoding the 14-3-3 epsilon isoform from Paracentrotus lividus sea urchin embryos, referred to as Pl14-3-3ε, and the phylogenetic relationship with other homologues described in different phyla. Pl14-3-3ε mRNA levels were measured by QPCR during development and found to increase from the mesenchyme blastula to the prism stage. In response to UV-B (312 nm) exposure, early stage embryos collected 2 h later showed a 2.3-fold (at 400 J/m(2)) and a 2.7-fold (at 800 J/m(2)) increase in Pl14-3-3ε transcript levels compared with controls. The spatial expression of Pl14-3-3ε mRNA, detected by whole mount in situ hybridization in both control and UV-B exposed embryos, harvested at late developmental stages, showed transcripts to be located in the archenteron of gastrula stage and widely distributed in all germ layers, respectively. The Pl14-3-3ε mRNA delocalization parallels the failure in archenteron elongation observed morphologically, as well as the lack of specific endoderm markers, investigated by indirect immuno-fluorescence on whole mount embryos. Results confirm the involvement of 14-3-3ε in the stress response elicited by UV-B and demonstrate, for the first time, its contribution at the transcriptional level in the sea urchin embryo.
Russo
Roberta
R
Consiglio Nazionale delle Ricerche, Istituto di Biomedicina e Immunologia Molecolare, Alberto Monroy, Via Ugo La Malfa 153, 90146, Palermo, Italy.
Zito
Francesca
F
Costa
Caterina
C
Bonaventura
Rosa
R
Matranga
Valeria
V
D048948:14-3-3 Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D004625:Embryo, Nonmammalian / Q000378:metabolism* / Q000528:radiation effects; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000378:metabolism; D012617:Sea Urchins / Q000196:embryology / Q000378:metabolism / Q000528:radiation effects; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D014466:Ultraviolet Rays
D048948:14-3-3 Proteins; D012333:RNA, Messenger