1. Archipelago/ hCdc4 By: Joe Pietropaoli

2. Review: Ubiquitylation E1 – ubiquitin-activating enzyme E2 – ubiquitin-conjugating enzyme E3 – ubiquitin-protein ligase www.myoops.org/.../0/chp_ub_p_sys.jpg

3. CDC 4 *Specific type of E3 ligase F-box Protein 7 WD40 protein-binding motifs Rajagopalan et al. Fig. 1-a

4. CDC 4 as E3 Ligase Ariizumi et al. Fig. 2

5. CDC 4 First Discovered in Budding Yeast Pintard et al. Fig. 1

6. CDC 4 Homologues Drosophila: Archipelago (Ago) Mammals: Cdc 4 (humans = hcdc 4) Both target cyclin E in respective organisms

7. Ago in Drosophila Development Growth (Wild-type embryos stained (in vivo evidence of with Ago Specific Anti Serum) association with dmyc) Moberg et al. Fig. 5-aMortimer et al. Fig. 1

8. CDC 4 in Mammals Knockout Mouse Experiment: homozygous mutant mice (cdc4-/-) -Subjects dies at about 10 days of Embryogenesis -Mice suffered from intracranial hemorrhaging -Mice did not show variation of cyclin E, and maintained cyclin E controlled Cdk2 activity http://dailyscreenshots.de/screenshot/mike-tysons-punch-out/nes-1873781 Keiko et al.

9. CDC 4 in Mammals Knockout mouse Experiment (cont) heterozygous mutant mice (cdc4+/-) -viable; greater susceptibility to radiation induced tumorogenisis -most tumors maintained and expresses wt cdc4 allele -cdc4 = haploinsufficient tumor suppressor gene

10. CDC 4 in Mammals cont. In vitro trial knocking out hcdc 4 in human cell lines Rajagopalan et al. Fig. 2-a

11. Results: hCDC4-/- cells proved to be chromosomally unstable Situation Summary: Mutation and loss of function in hcdc4 does not allow the SCF molecule to assemble and associate with cyclin E. Resulting over expression of cyclin E causes CI. Rajagopalan et al. Fig. 3

12. Endometrial Cancer -Cancer of the Uterine Wall -Caused be somatic mutations followed by LOH of the wild-type allele -Consistent with Knudson’s two hit hypothesis of tumor suppressor genes. -No evidence of hereditary cases -Highly Curable http://www.instablogsimages.com/images/2007/09/13/alcohol-may-bump-risk-of-cancer-of-womb_9.jpg

13. Connecting hCDC4 and Endometrial Cancer a Two of these tumors also contained phos-cyclin E. Tumor grade (G3/G2)hCDC4 gene mutation Cyclin E protein (8)7/13/8 a Phos-cyclin E (7)6/16/7 Neither (38)16/221/15 Spruck et al. Table 2

14. Connecting hCDC4 to Endometrial Cancer No.No. MutationMutation CodonaCodona ResultResult LOH2LOH2 6GAG TATGAG TAT 124124 Glu TyrGlu Tyr + 7AAA (TC)A (2-bp insertion)AAA (TC)A (2-bp insertion) 371371 Lys Ser, term codon 376Lys Ser, term codon 376 + 8CGA TGACGA TGA 367367 Arg TerArg Ter + 9CGT CATCGT CAT 465465 Arg HisArg His nibnib 1010 CGA CAACGA CAA 479479 Arg GlnArg Gln + 1 CGA TGACGA TGA 658658 Arg TerArg Ter + 1212 AGA A(A)G (1-bp insertion)AGA A(A)G (1-bp insertion) 472472 Arg Lys, term codon 476Arg Lys, term codon 476 + 1414 GTT ATTGTT ATT 23c23c Val IleVal Ile + No.MutationCodon a ResultLOH 4q32 6GAG TAT124Glu Tyr+ 7AAA (TC)A (2-bp insertion)371Lys Ser, term codon 376+ 8CGA TGA367Arg Ter+ 9CGT CAT465Arg Hisni b 10CGA CAA479Arg Gln+ 11CGA TGA658Arg Ter+ 12AGA A(A)G (1-bp insertion)472Arg Lys, term codon 476+ 14GTT ATT23 c Val Ile+ Spruck et al. Table 3

15. Endometrial Cancer: Diagnosis Risk Factors taking tamoxifen taking estrogen alone Diagnosis first detected by a pelvic screen confirmed by endometrial biopsy

16. Standard Treatments Surgery Chemotherapy Radiation Therapy Hormone Therapy

17. Clinical Trial Study Shows In advanced endometrial cancer, chemotherapy is better than whole- abdominal radiation Chemotherapy comes after an initial operation Chemotherapy is a combination of drugs: doxorubicin (adriamycin ® ) and cysplatin (platinol ® )