1. BC212/week7/20031 BC212 Signal transduction & growth control Death receptor signaling

2. BC212/week7/20032 Lecture overview Death receptor signaling pathways: TNF/TNFR gene-families DISC signaling NF-kappaB signaling ------------------------------------------ Discussion: FLIP: a switch between death & survival?

3. BC212/week7/20033 Lecture overview Death receptor signaling pathways: TNF/TNFR gene-families

4. BC212/week7/20034 TNF/TNFR families Ligands = typically membrane bound; at least 18 Receptors = one subunit; at least 28 Receptor-ligand promiscuity Decoy receptors Trimerization PLAD: pre-ligand assembly domain

5. BC212/week7/20035 TNF/TNFR families Ashkenazi Nat Rev Cancer 2:420, 2002

6. BC212/week7/20036 TNF/TNFR families

7. BC212/week7/20037 TNF/TNFR families Ashkenazi Nat Rev Cancer 2:420, 2002

8. BC212/week7/20038 TNF/TNFR families Goldstein Science 288:2388, 2000

9. BC212/week7/20039 TNF/TNFR families PLAD: pre-ligand assembly domain ALS (autoimmune lymphoproliferative syndrome): del 52-96 (het) Siegel et al. Science 288:2354, 2000

10. BC212/week7/200310 Lecture overview

11. BC212/week7/200311 DISC signaling Cell death, survival and cancer Intrinsic versus extrinsic death pathways

12. BC212/week7/200312 DISC signaling Lost track of this reference

13. BC212/week7/200313 DISC signaling Death receptor ligands TRAIL (DR4, DR5) FasL (Fas) TNF (TNFR1) TWEAK (TWEAK-R)

14. BC212/week7/200314 DISC signaling

15. BC212/week7/200315 DISC signaling Death receptor pathway components: FADD DD & DED Caspase-8, -10 DED & caspase (protease) FLIP Dominant inhibitor caspase relative Caspase-3 (etc.) Extrinsic pathway (via Bid)

16. BC212/week7/200316 DISC signaling Ashkenazi Nat Rev Cancer 2:420, 2002

17. BC212/week7/200317 DISC signaling

18. BC212/week7/200318 DISC signaling

19. BC212/week7/200319 DISC signaling Identifying components Fas screen for cytotoxic antibodies antibody crosslinks/multimerizes Fas FasL cell line which killed Fas-expressing cells (only) FasFc blocked killing Used biotinylated FasFc to affinity purify FasL FADD two-hybrid screen for DD interactors

20. BC212/week7/200320 DISC signaling lpr and gld mice Lymphoproliferative phenotype Resembles SLE lpr: homozygous mutation in Fas DD gld: homozygous inactivating mutation in FasL Implicates Fas-FasL in immune cell apoptosis

21. BC212/week7/200321 DISC signaling Ashkenazi Nat Rev Cancer 2:420, 2002

22. BC212/week7/200322 DISC signaling Induced proximity model of caspase activation Parallels with kinase activation Evidence: i) artificial dimerization activates caspase ii) zymogen has low, but detectable, activity

23. BC212/week7/200323 Lecture overview

24. BC212/week7/200324 NFkappaB signaling Haefner Drug Discovery Today 7:653 2002

25. BC212/week7/200325 NFkappaB signaling Karin, et al Nat Rev Cancer 2:301, 2002

26. BC212/week7/200326 NFkappaB signaling Haefner Drug Discovery Today 7:653 2002

27. BC212/week7/200327 NFkappaB signaling Karin, et al Nat Rev Cancer 2:301, 2002

28. BC212/week7/200328 NFkappaB signaling Karin, et al Nat Rev Cancer 2:301, 2002

29. BC212/week7/200329 NFkappaB signaling Evidence linking NFkappaB to proliferation/cancer RelA: viral NFkappaB homolog incrd stability; decrd susceptibility to degradation Bcl3: chromosomal translocations in leukemia over-expression NFKB2: chromosomal translocation in leukemia/lymphoma deletes IkappaB part

30. BC212/week7/200330 NFkappaB signaling Evidence establishing mechanism of NFkappaB signaling 1.Isolation via enhancer affinity chromatography 2.PMA (cells) & DOC (extracts) induces EMSA activity 3.PMA stimulates transport to the nucleus 4.DOC-dissociable inhibitor (IkappaB) 5.NFkB-IkB complex disrupted, EMSA induced, by PKC 6.IkB degradation: proteosome inhs, Ub’n, etc.

31. BC212/week7/200331 NFkappaB signaling

32. BC212/week7/200332 NFkappaB signaling

33. BC212/week7/200333 Lecture overview

34. BC212/week7/200334 FLIP as a switch?