Ectoderm nuclei from sea urchin embryos contain a Spec-DNA binding protein similar to the vertebrate transcription factor USF.
The Spec gene family of Stronglyocentrotus purpuratus is expressed exclusively in aboral ectoderm cells during embryogenesis. To investigate the regulation of Spec gene activity, the region around the Spec1 transcriptional initiation site was analyzed for sites of protein-DNA interaction. One high-affinity site bound a factor termed SpF1 within the Spec1 5' untranslated leader region at position +39 to +60. The core sequence recognized by SpF1, CACGTG, is the same as that of the upstream stimulatory factor (USF), a widely occurring vertebrate transcription factor containing a myc-HLH motif. A comparison of USF- and SpF1-binding activities suggested that SpF1 was a sea urchin version of USF. SpF1 activity was detectable only in ectoderm cells of the embryo, implying that it has a role as a cell type-specific transcription factor. SpF1-binding sites were also found upstream of the Spec2a and Spec2c genes in the same conserved sequence block as Spec1. Extracts from Lytechinus pictus embryos showed an SpF1-like activity, suggesting that SpF1 is conserved in sea urchins. Surprisingly, changes in the Spec1, Spec2a, or Spec2c genes that removed or modified the SpF1-binding site had no effect on expression when reporter gene fusions containing these mutations were injected into sea urchin eggs and analyzed for expression during embryogenesis. We propose that, while SpF1 may not be essential for expression of the exogenously introduced reporter genes, it may be required for proper regulation of the endogenous Spec genes.
Tomlinson
C R
CR
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Kozlowski
M T
MT
Klein
W H
WH
D000818:Animals; D002467:Cell Nucleus / Q000502:physiology*; D004268:DNA-Binding Proteins; D004475:Ectoderm / Q000502:physiology*; D005821:Genetic Techniques; D012617:Sea Urchins / Q000196:embryology* / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*; D051720:Upstream Stimulatory Factors
D004268:DNA-Binding Proteins; D014157:Transcription Factors; D051720:Upstream Stimulatory Factors