title: An Elk transcription factor is required for Runx-dependent survival signaling in the sea urchin embryo.
abstract: Elk proteins are Ets family transcription factors that regulate cell proliferation, survival, and differentiation in response to ERK (extracellular-signal regulated kinase)-mediated phosphorylation. Here we report the embryonic expression and function of Sp-Elk, the single Elk gene of the sea urchin Strongylocentrotus purpuratus. Sp-Elk is zygotically expressed throughout the embryo beginning at late cleavage stage, with peak expression occurring at blastula stage. Morpholino antisense-mediated knockdown of Sp-Elk causes blastula-stage developmental arrest and embryo disintegration due to apoptosis, a phenotype that is rescued by wild-type Elk mRNA. Development is also rescued by Elk mRNA encoding a serine to aspartic acid substitution (S402D) that mimics ERK-mediated phosphorylation of a conserved site that enhances DNA binding, but not by Elk mRNA encoding an alanine substitution at the same site (S402A). This demonstrates both that the apoptotic phenotype of the morphants is specifically caused by Elk depletion, and that phosphorylation of serine 402 of Sp-Elk is critical for its anti-apoptotic function. Knockdown of Sp-Elk results in under-expression of several regulatory genes involved in cell fate specification, cell cycle control, and survival signaling, including the transcriptional regulator Sp-Runt-1 and its target Sp-PKC1, both of which were shown previously to be required for cell survival during embryogenesis. Both Sp-Runt-1 and Sp-PKC1 have sequences upstream of their transcription start sites that specifically bind Sp-Elk. These results indicate that Sp-Elk is the signal-dependent activator of a feed-forward gene regulatory circuit, consisting also of Sp-Runt-1 and Sp-PKC1, which actively suppresses apoptosis in the early embryo.
authors.affiliation: Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, Napoli 80121, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Rizzo
authors.affiliation: MDI Biological Laboratory, Salisbury Cove, ME 04672, USA.
authors.forename: James A
authors.initials: JA
authors.lastname: Coffman
authors.affiliation: Biology and Evolution of Marine Organisms, Stazione Zoologica Anton Dohrn, Napoli 80121, Italy. Electronic address: miarnone@szn.it.
authors.forename: Maria Ina
authors.initials: MI
authors.lastname: Arnone
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D036703:Blastula; D002470:Cell Survival / Q000235:genetics; D050657:Core Binding Factor alpha Subunits / Q000378:metabolism*; D047108:Embryonic Development; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D016376:Oligonucleotides, Antisense; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D012617:Sea Urchins / Q000196:embryology* / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics; D050801:Ternary Complex Factors / Q000378:metabolism*
chemical_list: D050657:Core Binding Factor alpha Subunits; D016376:Oligonucleotides, Antisense; D050801:Ternary Complex Factors
keywords: Apoptosis; ERK; Elk; Embryo; PKC; Runx; Sea urchin