Altering cell fates in sea urchin embryos by overexpressing SpOtx, an orthodenticle-related protein.
While many general features of cell fate specification in the sea urchin embryo are understood, specific factors associated with these events remain unidentified. SpOtx, an orthodenticle-related protein, has been implicated as a transcriptional activator of the aboral ectoderm-specific Spec2a gene. Here, we present evidence that SpOtx has the potential to alter cell fates. SpOtx was found in the cytoplasm of early cleavage stage embryos and was translocated into nuclei between the 60- and 120-cell stage, coincident with Spec gene activation. Eggs injected with SpOtx mRNA developed into epithelial balls of aboral ectoderm suggesting that SpOtx redirected nonaboral ectoderm cells to an aboral ectoderm fate. At least three distinct domains on SpOtx, the homeobox and regions in the N-terminal and C-terminal halves of the protein, were required for the morphological alterations. These same N-terminal and C-terminal regions were shown to be transactivation domains in a yeast transactivation assay, indicating that the biological effects of overexpressing SpOtx were due to its action as a transcription factor. Our results suggest that SpOtx is involved in aboral ectoderm differentiation by activating aboral ectoderm-specific genes and that modulating its expression can lead to changes in cell fate.
Mao
C A
CA
Department of Biochemistry and Molecular Biology, The University of Texas MD Anderson Cancer Center, Houston 77030, USA.
Wikramanayake
A H
AH
Gan
L
L
Chuang
C K
CK
Summers
R G
RG
Klein
W H
WH
D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002451:Cell Compartmentation; D002452:Cell Count; D002467:Cell Nucleus / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D029721:Drosophila Proteins; D004475:Ectoderm / Q000166:cytology; D005455:Fluorescent Antibody Technique; D005801:Genes, Homeobox; D018398:Homeodomain Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D012313:RNA; D012617:Sea Urchins / Q000196:embryology* / Q000235:genetics; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic
D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; C039919:RNA, recombinant; C092316:SpOtx protein, Strongylocentrotus purpuratus; C093053:oc protein, Drosophila; D012313:RNA