Regulation of HP1–chromatin binding by histone H3 methylation and phosphorylation

Tri-methylation of histone H3 lysine 9 is important for recruiting heterochromatin protein 1 (HP1) to discrete regions of the genome, thereby regulating gene expression, chromatin packaging and heterochromatin formation. Here we show that HP1α, -β, and -γ are released from chromatin during the M phase of the cell cycle, even though tri-methylation levels of histone H3 lysine 9 remain unchanged. However, the additional, transient modification of histone H3 by phosphorylation of serine 10 next to the more stable methyl-lysine 9 mark is sufficient to eject HP1 proteins from their binding sites. Inhibition or depletion of the mitotic kinase Aurora B, which phosphorylates serine 10 on histone H3, causes retention of HP1 proteins on mitotic chromosomes, suggesting that H3 serine 10 phosphorylation is necessary for the dissociation of HP1 from chromatin in M phase. These findings establish a regulatory mechanism of protein–protein interactions, through a combinatorial readout of two adjacent post-translational modifications: a stable methylation and a dynamic phosphorylation mark.

[1]  D. Moazed,et al.  Heterochromatin and Epigenetic Control of Gene Expression , 2003, Science.

[2]  T. Kapoor,et al.  Correcting improper chromosome–spindle attachments during cell division , 2004, Nature Cell Biology.

[3]  Andrew J. Bannister,et al.  Selective recognition of methylated lysine 9 on histone H3 by the HP1 chromo domain , 2001, Nature.

[4]  S. Khorasanizadeh.  The Nucleosome From Genomic Organization to Genomic Regulation , 2004, Cell.

[5]  H. Saya,et al.  Autophosphorylation of a Newly Identified Site of Aurora-B Is Indispensable for Cytokinesis* , 2004, Journal of Biological Chemistry.

[6]  Sean D. Taverna,et al.  Specificity of the HP1 chromo domain for the methylated N‐terminus of histone H3 , 2001, The EMBO journal.

[7]  J. Swedlow,et al.  Mitotic mechanics: the auroras come into view. , 2003, Current opinion in cell biology.

[8]  S. Diekmann,et al.  High- and low-mobility populations of HP1 in heterochromatin of mammalian cells. , 2004, Molecular biology of the cell.

[9]  X. Liu,et al.  Survivin Enhances Aurora-B Kinase Activity and Localizes Aurora-B in Human Cells* , 2003, The Journal of Biological Chemistry.

[10]  D. Kioussis,et al.  Modulation of Heterochromatin Protein 1 Dynamics in Primary Mammalian Cells , 2003, Science.

[11]  Danny Reinberg,et al.  Histone lysine methylation: a signature for chromatin function. , 2003, Trends in genetics : TIG.

[12]  E. Nigg,et al.  Exploring the functional interactions between Aurora B, INCENP, and survivin in mitosis. , 2003, Molecular biology of the cell.

[13]  R. Allshire,et al.  Kinetochore and heterochromatin domains of the fission yeast centromere , 2004, Chromosome Research.

[14]  S. Elgin,et al.  The HP1 protein family: getting a grip on chromatin. , 2000, Current opinion in genetics & development.

[15]  C. Ponting,et al.  Regulation of chromatin structure by site-specific histone H3 methyltransferases , 2000, Nature.

[16]  Y. Hiraoka,et al.  Cell cycle behavior of human HP1 subtypes: distinct molecular domains of HP1 are required for their centromeric localization during interphase and metaphase , 2003, Journal of Cell Science.

[17]  Stephen S. Taylor,et al.  The kinase haspin is required for mitotic histone H3 Thr 3 phosphorylation and normal metaphase chromosome alignment. , 2005, Genes & development.

[18]  Zu-Wen Sun,et al.  Mitotic Phosphorylation of Histone H3 Is Governed by Ipl1/aurora Kinase and Glc7/PP1 Phosphatase in Budding Yeast and Nematodes , 2000, Cell.

[19]  D. Kirschmann,et al.  Does heterochromatin protein 1 always follow code? , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[20]  A. Murray,et al.  Cell cycle extracts. , 1991, Methods in cell biology.

[21]  M. Groudine,et al.  Controlling the double helix , 2003, Nature.

[22]  Karl Mechtler,et al.  Methylation of histone H3 lysine 9 creates a binding site for HP1 proteins , 2001, Nature.

[23]  S. Grewal,et al.  Heterochromatin: new possibilities for the inheritance of structure. , 2002, Current opinion in genetics & development.

[24]  J. Martens,et al.  Partitioning and plasticity of repressive histone methylation states in mammalian chromatin. , 2003, Molecular cell.

[25]  Patrick England,et al.  Tethering of HP1 proteins to chromatin is relieved by phosphoacetylation of histone H3 , 2004, EMBO reports.

[26]  J. Peters,et al.  The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore–microtubule attachment and in maintaining the spindle assembly checkpoint , 2003, The Journal of cell biology.

[27]  G. Schotta,et al.  The indexing potential of histone lysine methylation. , 2004, Novartis Foundation symposium.

[28]  S. Dimitrov,et al.  Phosphorylation of serine 10 in histone H3, what for? , 2003, Journal of Cell Science.

[29]  T. Jenuwein,et al.  Higher-order structure in pericentric heterochromatin involves a distinct pattern of histone modification and an RNA component , 2002, Nature Genetics.

[30]  Wolfgang Fischle,et al.  Binary switches and modification cassettes in histone biology and beyond , 2003, Nature.

[31]  M. Grunstein,et al.  Centromere Silencing and Function in Fission Yeast Is Governed by the Amino Terminus of Histone H3 , 2003, Current Biology.

[32]  Tom Misteli,et al.  Maintenance of Stable Heterochromatin Domains by Dynamic HP1 Binding , 2003, Science.

[33]  Youngchang Kim,et al.  Molecular basis for the discrimination of repressive methyl-lysine marks in histone H3 by Polycomb and HP1 chromodomains. , 2003, Genes & development.

[34]  Stephen S. Taylor,et al.  Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores , 2003, The Journal of cell biology.

[35]  G. Almouzni,et al.  HP1 and the dynamics of heterochromatin maintenance , 2004, Nature Reviews Molecular Cell Biology.

[36]  Ernest D Laue,et al.  Structural basis of HP1/PXVXL motif peptide interactions and HP1 localisation to heterochromatin , 2004, The EMBO journal.

[37]  S. Jacobs,et al.  Structure of HP1 Chromodomain Bound to a Lysine 9-Methylated Histone H3 Tail , 2002, Science.

[38]  Ryoma Ohi,et al.  The Chromosomal Passenger Complex Is Required for Chromatin-Induced Microtubule Stabilization and Spindle Assembly , 2004, Cell.

[39]  Alexey G. Murzin,et al.  Structure of the HP1 chromodomain bound to histone H3 methylated at lysine 9 , 2002, Nature.

[40]  Y. Allory,et al.  Localization and phosphorylation of HP1 proteins during the cell cycle in mammalian cells , 1999, Chromosoma.

[41]  Scott A. Busby,et al.  Novel linear quadrupole ion trap/FT mass spectrometer: performance characterization and use in the comparative analysis of histone H3 post-translational modifications. , 2004, Journal of proteome research.

[42]  K. Sugimoto,et al.  Molecular behavior in living mitotic cells of human centromere heterochromatin protein HPLalpha ectopically expressed as a fusion to red fluorescent protein. , 2001, Cell structure and function.

[43]  J. Wong,et al.  Relationship between Histone H3 Lysine 9 Methylation, Transcription Repression, and Heterochromatin Protein 1 Recruitment , 2005, Molecular and Cellular Biology.

[44]  Andrew W. Murray,et al.  Chapter 30 Cell Cycle Extracts , 1991 .

[45]  C. Allis,et al.  Mitosis-specific phosphorylation of histone H3 initiates primarily within pericentromeric heterochromatin during G2 and spreads in an ordered fashion coincident with mitotic chromosome condensation , 1997, Chromosoma.

[46]  S. Jacobs,et al.  Assays for the determination of structure and dynamics of the interaction of the chromodomain with histone peptides. , 2004, Methods in enzymology.

[47]  C. Allis,et al.  Phosphorylation of Histone H3 Is Required for Proper Chromosome Condensation and Segregation , 1999, Cell.

[48]  Osamu Iwasaki,et al.  A conserved Mis12 centromere complex is linked to heterochromatic HP1 and outer kinetochore protein Zwint-1 , 2004, Nature Cell Biology.