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P01 Videoconference
1. Andrew M. Scharenberg M.D. Children’s Hospital and Regional Medical Center, Seattle, WA
2. Feske, Nature Reviews in Immunology 2007 Fox, Hammerman, and Thompson, Nature Reviews Immunology 2005 Proliferation Quiescence
3. Channel domain: gating regulated by [Mg 2+ ] : permeates divalent cations Kinase domain: phosphotransferase activity not required for channel gating, substrates still under investigation amino terminal unique region transmembrane Kinase domain CCR TRPM7 TRPM7-conditional KO tamoxifen-regulated kb 2.0 1.0 P KO KO Targeting construct II (HisD) B B His D probe 5’ flank 3’ flank Targeted region Targeting construct I (neomycin-loxP) loxP loxP Model system: DT40 chicken B-cell line Kb 1 2 3 4 ‘ 5 6 TM
4. parent KO +10 Mg 2+ KO TRPM7 TRPM6 amino terminal unique region transmembrane Kinase domain CCR CCR Targeting construct II (HisD) Kb 1 2 3 4 ‘ 5 6 TM B B His D Targeting construct I (neomycin) B B NeoR KO KO + 10 Mg Cells/ml x 10^6 Days 0 1 2 3 0 1 2 3 4 5 WT Cl1 Cl2 Cl3 par ctl TRPM7
5. TRPM6: organism TRPM7: cellular Channel Closed cell Mg 2+ replete Channel Open cell needs Mg 2+ Local increase in [Mg 2+ ] TRPM7 TRPM6 amino terminal unique region transmembrane Kinase domain CCR CCR 1.0 0.5 0 WT cWT WT/high low Tot. Mg reg media/ tot Mg supp media Cells/ml x 10^6 very low WT/high + 10 mM Mg 2+ cWT
6. Decreased rate of growth ( addition of mass) Cell size over the cell cycle G1 S G2 M Decreased rate of DNA synthesis G1 S G2 M Decreased rate of Mitosis G1 S G2 M x Increased rate of death G1 S G2 M
