2. Myc genes are deregulated in human cancers Myc gene deregulated Cancer type c-myc Bladder, breast, colon, gastric, melanoma, myeloma, ovary, prostate, lung N-myc Neuroblastoma, breast, lung L-myc Breast, lung
3. Myc target genes - Myc target genes are involved in proliferation, vasculogenesis, cell adhesion RNA POL I ribosomal RNA Myc RNA POL II Growth metabolism, translation, ribosomal proteins RNA POL III tRNAs
4. Myc expression induces cell growth Control dMyc (GMM) Gal4 GMR Gal4 dMyc Gal4 UAS dMyc dMyc dMyc Expression of dMyc in post-mitotic cells during eye development GMR-Gal4, UAS-dMyc
14. Lid is required for dMyc to activate Nop60B ? Lid Nop60B E-box
15. JmjC is a lysine demethylase domain Nature, 2006 - JHDM1A demethylates histone H3 lysine 36 - Enzymatic activity requires the JmjC domain, Fe2+ and a-ketoglutarate
16. M M M M M M CH3 CH3 CH3 H3C CH3 CH3 trimethyl dimethyl monomethyl unmodified N+ NH+ NH2+ NH3+ JmjC-dependent demethylase activities H3 ARTKYTARKSTGGKAPRKQLATKAARKSAPATGGVK..K..K H4 SGRGKGGKGLGKGGAKRHRKVLR
17. What is the mechanism of Lid function? JmjC PHD PHD PHD ARID JmjN C5HC2 Is Lid a histone demethylase? Is this activity required for dMyc-induced cell growth?
19. Histone H3 lysine 4 methylation correlates with transcriptional activation Correlation with transcription + K4me3 + K4me2 +/- K4me1 Adapted from Li and Workman, 2007 - Lid’s demethylase activity implicates it as a transcriptional repressor
20. * * dMyc Lid-JmjC* Lid’s JmjC-encoded demethylase activity is not required for dMyc function JmjC PHD PHD PHD ARID JmjN C5HC2 NLS dMyc Lid dMyc - Expression of either wildtype or JmjC* Lid alone has no effect on eye development
21. Myc What is the mechanism of Lid function? ? Lid Nop60B E-box
22. Enhanced? dMyc Lid dMyc dMyc Lid-JmjC* dMyc Lid-D Identifying domains of Lid required for Myc-induced cell growth? Lid domain UAS
23. Domains of Lid required for dMyc function Enhance dMyc phenotype? JmjN JmjC PHD PHD PHD ARID C5HC2 NLS * * X ( ) ( ) ( ) X ( ) ( ) nd X
24. TFIID TAF3 H3K4me3 Transcriptional initiation PHD fingers read the histone code Nurf BPTF Pygo BHC80 H3K4me0 H3K4me2 H3K4me2/3 Wnt-induced transcriptional activation Tethers the LSD1 repressor to chromatin Nucleosome remodeling
26. c-Myc binding correlates with high levels of di and trimethylated H3K4 - Histone H3 lysine 4 methylation precedes and is independent of Myc binding.
27.
28. One of the functions on Lid might be to ‘recruit’ Myc to sites that are enriched for H3K4me2/3
29. Thank you Einstein Christina Greer FHCRC Ling Li Bob Eisenman Salk institute Satchin Panda Hiep Le