1. Conserve Fibroblast growth factor 8 (fgf 8) domains Ana Tomas Judith Paridaen Susana Domingues

2. Fibroblast growth factor 8 Member of the FGF family Function: Mitogenic Chemotactic Angiogenic activity Promoting cell growth, differentiation and motility

3. Some Fgf 8 characteristics N.º of a.a.: 233 Molecular weight: 26525.4 Theoretical p.i.: 10.44 Total n.º of negatively charged residues (Asp + Glu): 19 Total n.º of positively charged residues (Arg + Lys): 40 Formula: C 1164 H 1877 N 363 O 330 S 9 Total n.º of atoms: 3743 Grand average of hydropathicity: -0.651 (soluble) Estimated half-life: The N-terminal of the sequence considered is M (Met) The estimated half-life is:  30 hours (mammalian) Instability index: The instability index (II) is computed to be 47.80 Unstable protein ProtParam tool - ExPASy

4. Structure 6 chains present 913 residues

5. Structure and active sites HBGF/FGF SP binding ProSAT - Protein Structure Annotation Tool

6. Models Axolotl Chicken Ciona S. Cow Dog Drosophila Fugu Houseshrew Human Japanese lamprey Mouse Opossum Oryzias Rabbit Rat Rhesus macaque Shell turtle Tetraodon Zebrafish Ciona

7. Phylogenetic tree Cluster algorithm

8. Consensus domains p85_SH2 SH2 KGAETGFYICMNKRG InsR_Kin Y_kin NARYEGWYMAFTRRG ErkDD Kin_bind RDCIFTEIVLENNYT Multalin – Multi alignment program

9. 3D structure and consensus domains ErkDD Kin_bind RDCIFTEIVLENNYT InsR_Kin Y_kin NARYEGWYMAFTRRG p85_SH2 SH2 KGAETGFYIC MNKRG Rasmol

10. Fgf8 structure with receptor Rasmol Fgf Receptor

11. Common regulatory elements in Fgf8 downstream targets Anna Ferrer-Vaquer Alisa Fuchs

12. Fgf8 signaling pathway  Elk  Ets  cJun  Fos  Myc ...

13. Fgf8 inducible genes Sprouty1 Pax3 Otx2 Tbx2 Erm Pea3 Ptx1 Lhx6 Sef AP2 Flrt3.... Many more Sprouty1 > Mm, Gg, Dr Pax3 > Mm, Gg, Dr Otx2 > Mm, Gg, Dr Tbx2 > Mm, Gg, Dr Erm Pea3 Ptx1 Lhx6 Sef AP2 Flrt3.... Many more  Limitation:  1Kb of 5’UTR

14. Alignment Alignment of sequences from different genes to find consensus blocks Not reliable

15. Promoter analysis

16. Results of promoter analysis Promotor/TFElkvMybZBP-89 mSprouty1 +++ mPax3 + mOtx2 +++ mTbx2 +++

17. Conclusion Analysis of nucleotide sequences is much more difficult than amino acids