DEV014282 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Structure and phylogenetic relationships of sea urchin FGFA and FGFR2. (A) Domain structure of the deduced FGFA protein sequence. (B) Amino acid alignment of the core FGF domain of the predicted sea urchin (P. lividus and S. purpuratus) FGFA protein with FGF9, FGF16 or FGF20 related proteins from human (Hs), mouse (Mm) and zebrafish (Dr). Dark highlighting indicates positions where at least 50% of the residues are identical. Amino acids highly conserved among FGFs are marked with a red asterisk, whereas amino acids implicated in heparin binding are marked with a black asterisk. The position of the 12 β-strands that constitute the FGF trefoil is indicated under the alignment. The positions of the two introns are indicated by arrowheads and the positions of the PCR primers used to characterize the splicing products generated in the presence of the splice inhibiting morpholino are indicated in red. (C) Phylogenetic relationships of sea urchin FGFA with different FGF-family members from zebrafish and Drosophila using the Maximum likelihood method. The numbers above the nodes represent bootstrap values in % (500 iterations). (D) Genomic organization of the fgfr2 locus based on the Spur_v2.1 release. The gene prediction programs used to annotate the genome sequence of Strongylocentrotus had failed to predict a receptor tyrosine kinase at the FGFR2 locus. They had instead predicted two separate gene models, one encoding a protein with a tyrosine kinase domain (CG10244) and another encoding a protein with immunoglobulin-like domains related to N-CAM (hmm 93681). Comparison with the FGFR2 cDNA shows that the two predictions are different parts of the same gene. (E) Domain structure of FGFR2. The conserved Ig-like (immunoglobulin-like) domains are shown in orange, the FN3 (fibronectin 3) in light blue, transmembrane region and the Tyrosine kinase domain in dark blue. (F) Sequence alignments between FGFR2 and FGFR1 from sea urchin and the sequences giving the highest BLAST scores against FGFR2. Spodoptera frugiperda (Sf), Bombyx mori (Bm), and Branchiostoma belcheri (Bb). Phylogenetic relationships between FGFR2 and other RTKs and FGFRs from vertebrates and invertebrates.

• Supplemental Figure S2 -

  Fig. S2. Injection of a control morpholino (1 mM) directed against the Nemo-like kinase transcript does not interfere with gastrulation or with patterning of the skeleton. vv, vegetal view.