Histone Modification.BCH6415.Spr2006.Print Ver Part I

Histone Modification.BCH6415.Spr2006.Print Ver Part I -...

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HISTONE MODIFICATION BCH 6415 Spring 2006 (Part I)
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Schematic of ChIP Immuno- preciptation with specific AB Chromatin purification Reverse of Crosslinking DNA purification PCR of genomic fragments 500 bp Sonication Y K562 Input No AB α USF II TF II I NF-E2 with formaldehyde Crosslinking
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ADP-ribosylation Sumoylation Biotinylation Citrulline…. .
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Covalent Modifications of Histone Tails Lusser Curr Opin Plant Biol 5:437;2002 Turner, BM Cell 2002
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Fr. Peterson & Laniel Curr Biol. 2005
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Histone Acetylation/Deacetylation Known acetylation sites in core histone N-term. tails K = lysine Roth, et al. Ann Rev. Biochem. 70:81; 2001 Kuo & Allis Bioessays 1998 Turner, BM 1999 Wade, PA Hum Mol Genet. 2001
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Acetylation sites of various HAT’s Mamorstein & Roth Curr Op Genet. Devel . 2001 (see Yang, XJ NAR 2004 for updated list) HAT Families Two types of HAT’s: Type A – localized in nucleus; acetylates nucleosomal histones Type B – localized in cytoplasm; probably acetylates newly synthesized histones prior to chromatin assembly Five Familes of HATs: GNAT – show seq similarity to Gcn5 HAT MYST p300/CBP GTF HAT’s (e.g., TAF250) Nuclear hormone-related (e.g., SCR1, ACTR) Most HAT’s multisubunit complexes Some HAT’s can acetylate other proteins (e.g., TFIIE, TFIIF)
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Motifs in found in various HAT’s GNAT MYST Green – bromodomains Orange – chromodomains Red – GNAT HAT domain Pink – MYST homology region; HAT Light Blue & Green – Zn fingers Fr. Carrozza et al. Trends Genet 2003 Fr. Marmorstein & Roth Curr Opin 2001
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Forsberg et al. PNAS 97:14494; 2000 Histone acetylation pattern across the mouse β-globin locus in DMSO-induced MEL cells maj and minor genes expressed) Distribution of histone acetylation across loci and genes
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Filled bars: p16 expressing cells Open bars: non-expressing cells Histone acetylation levels across the human p16 locus In yeast, H3K9/14 acetylation extends across coding region Fr. Liang et al. PNAS 2004 Conclusion: Highest H3k9/14Ac levels localized to 5’ region of genes in higher eukaryotes.
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How does histone acetylation “open” nucleosomes/chromatin? BUT,
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Histone Modification.BCH6415.Spr2006.Print Ver Part I -...

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