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Epigenetik: Von der Genomforschung zur Epigenomforschung

Epigenetik: Von der Genomforschung zur Epigenomforschung. 4362. 4355. Exploring the hierarchy of epigenetic mechanisms. Chromosome territories, chromatin fibers and domains, interchromatin compartment, topology of active and silent genes. Histone modifications and chromatin remodelling.

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Epigenetik: Von der Genomforschung zur Epigenomforschung

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  1. Epigenetik:Von der Genomforschung zur Epigenomforschung

  2. 4362

  3. 4355

  4. Exploring the hierarchy of epigenetic mechanisms Chromosome territories,chromatin fibers and domains, interchromatin compartment, topology of active and silent genes Histone modifications and chromatin remodelling DNA methylation Horn and Peterson, 2002

  5. 2050

  6. 0777 Metaphasechromomen aus einer diploiden, menschlichen Zelle in der Mitose

  7. 2531

  8. 2531

  9. 733

  10. 2520

  11. 1. Herstellung von Metaphase-Präparaten aus Vollblutkulturen Markieren der Sonde mit Reportermolekülen (Biotin, etc.) bzw. Fluoreszenzfarbstoffen 3. Denaturierung der Sonden- und Ziel-DNA Sonden Ziel-DNA 4. Hybridisierung der Sonde an die komplementären Ziel-DNA Sequenzen; bei indirekter Markierung anschließende Immunodetektion 5. Visualisierung der Fluoreszenz- signale Sonde Ziel-DNA Fluoreszenz in situ Hybridisierung (FISH) 2. Herstellung der DNA-Sonde

  12. 3D map of chromosome territory arrangements in postmitotic human fibroblast cell nuclei (G0) “true color“ image (RGB) 24 color painting of all chromosome territories in male diploid human fibroblast nucleus classification (M-FISH 3D-program) Bolzer et al, 2005

  13. 2192

  14. the DNA sequence levelthe textbook story • linear array of sequence elements • transcription units • promoters, enhancers, silencers • sequences guide the machineries for transcription

  15. X m m AACGAGATCGTTCGAA AACGAGATCGTTCGAA X HeinrichLeonhardt

  16. DNA Methylierung Heinrich Leonhardt

  17. Heinrich Leonhardt

  18. de novo methylation Heinrich Leonhardt

  19. Transcriptional repression through DNA methylation HeinrichLeonhardt

  20. the chromatin level histone modifications • methylation,acetylation, phosphorylation, ubiquitinylation chromatin remodelling • ATP-dependent shiftingof nucleosomes

  21. Modification sites at the N-termini of histone tails Modifications above an amino acid are linked to transcriptionally active chromatin Modifications below an amino acid are linked to transcriptionally inactive chromatin K= Lysin, R= Arginin, S= Serin, Note that methylation can exist in a mono-, di- or trimethylated state

  22. Currently used antibodies against histone modification sites Transcriptionally active (eu)chromatin Mono-methyl H3-K9 Mono-methyl H4-K20 Unknown (active chromatin?) Transcriptionally active (eu)chromatin Acetyl-H4-K8 Tri-methyl H3-K27 Facultative Heterochromatin Transcriptionally repressed and Constitutive (hetero)chromatin Tri-methyl H3-K9 Tri-methyl H4-K20 H3/H4-multi-methyl lysine (MML) sites Mostly repressed chromatin

  23. N Lysine in histone: O O C C C - g e P N+ d DNA a b C C O cell membrane C O O [3H]-Ac Acetyl-CoA HAT (Histone Acetyl- Transferase) HDAC (Histone DeACetylase) C O C - O CoA N Uncharged Acetyl-Lysine in histone C C C N C e C C C C O O Histone Acetylation Enzymes Jakob Waterborg WaterborgJ@umkc.edu

  24. + charged histone tail acetylated histone tail Function of Histone Acetylation 30 nm 11 nm HIGH in histone H1 Often low in histone H1 Jakob Waterborg WaterborgJ@umkc.edu

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