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1. Introduction
2. Molecular identification of the CRAC channel
3. Outline
4. T cells in the lymph node
5. T cell - dendritic cell interactions in lymph node
6. T-Cell / APC interaction dynamics
7. Ca and gene expression in T cells
8. How does calcium enter the cell
9. Ion channels in T lymphocytes
10. Ca signaling in T lymphocytes
11. CRAC channel and lymphocyte activation
12. Genes encoding channel components
13. CRAC current in Drosophila S2 cells
14. Screening for CRAC in Drosophila S2 cells
15. Suppression of Ca influx by Stim dsRNA
16. Patch clamp recordings confirm STIM expression
17. Suppression of CRAC current by Stim dsRNA
18. STIM1 suppression in Jurkat T cells
19. Effects of Stim (STIM1) supression
20. Two functions of Stim
21. Stim sequence and domains
22. Overexpression of STIM and EF hand mutants
23. Activation of CRAC channels in S2 cells
24. Overexpression of wild-type STIM1 in the ER
25. Re-distribution of STIM1 following store depletion
26. Hotspots of STIM1 following store depletion
27. Subcellular distribution of STIM1: conclusion
28. Dual functions of Stim
29. Genome-wide screen for CRAC
30. Genome-wide RNAi screen for SOC influx
31. Quantitative screening results
32. Top 10 Hits
33. RNAi knock-down of olf186-F gene
34. olf186-F conserved gene family
35. Proposed olf186-F membrane topology
36. Overexpression of olf186-F and olf186-F + Stim
37. Accelerating CRAC current development
38. Functional expression olf186-F + STIM as SOC
39. Giant CRAC currents induced by olf186-F + Stim
40. Amplifying CRAC current
41. Molecular identity of the CRAC channel
42. Mutants of olf186-F co-expressed with Stim
43. Mutation E180D Orai alters I-V shape
44. CRAC current upon choline substitution for Na
45. E180D Orai alters divalent cation selectivity
46. Calcium block of CRAC current in E180D Orai
47. Altered monovalent ion selectivity of E180D Orai
48. More alterations in E180D Orai
49. Charge-neutralizing Orai mutations in S1-S2
50. pA/pF of WT and mutants in S1-S2
51. Gadolinium block in WT and mutants
52. olf186-F is a bona fide ion channel
53. CRAC = olf186-F (Orai, CRACM)
54. How does the CRAC channel open?
55. Stim / olf186-F co-immunoprecipitation
56. Syntaxin 5 (and tsr) knockdown
57. SERCA knockdown
58. CRAC channel activation pathway
59. Several possibilities for activation
60. CRAC summary
61. Acknowledgments (1)
62. Acknowledgments (2)

Topics Covered

• Background on Calcium signaling and ion channels in the immune system, role of CRAC channels in lymphocyte activation
• Discovery of Stim and STIM1 by RNA interference screen in Drosophila S2 cells
• Stim as an ER Calcium sensor and as a messenger to the plasma membrane
• Genome-wide RNA interference screen and the discovery of Orai
• Amplifying CRAC current by co-expression of Stim and Orai
• Molecular identification of CRAC channels by altered ion selectivity in a mutant of Orai
• How does the CRAC channel activate?

Links

Series:

• Calcium Signaling I

Categories:

• Cell Biology
• Immunology

Therapeutic Areas:

• Immunology & Inflammation

Talk Citation

Publication History

• Published on October 1, 2007

Financial Disclosures

• Prof. Michael Cahalan has not informed HSTalks of any commercial/financial relationship that it is appropriate to disclose.