The ankyrin repeats of G9a and GLP histone methyltransferases are mono- and dimethyllysine binding modules

Histone modifications have important roles in transcriptional control, mitosis and heterochromatin formation. G9a and G9a-like protein (GLP) are euchromatin-associated methyltransferases that repress transcription by mono- and dimethylating histone H3 at Lys9 (H3K9). Here we demonstrate that the ankyrin repeat domains of G9a and GLP bind with strong preference to N-terminal H3 peptides containing mono- or dimethyl K9. X-ray crystallography revealed the basis for recognition of the methylated lysine by a partial hydrophobic cage with three tryptophans and one acidic residue. Substitution of key residues in the cage eliminated the H3 tail interaction. Hence, G9a and GLP contain a new type of methyllysine binding module (the ankyrin repeat domains) and are the first examples of protein (histone) methyltransferases harboring in a single polypeptide the activities that generate and read the same epigenetic mark.

[1]  M. Walsh,et al.  CCAAT displacement protein/cut homolog recruits G9a histone lysine methyltransferase to repress transcription. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[2]  Anjanabha Saha,et al.  ING2 PHD domain links histone H3 lysine 4 methylation to active gene repression , 2006, Nature.

[3]  M. Sternberg,et al.  Enhanced genome annotation using structural profiles in the program 3D-PSSM. , 2000, Journal of molecular biology.

[4]  Tsutomu Ohta,et al.  Histone methyltransferases G9a and GLP form heteromeric complexes and are both crucial for methylation of euchromatin at H3-K9. , 2005, Genes & development.

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

[6]  F. Studier,et al.  Protein production by auto-induction in high density shaking cultures. , 2005, Protein expression and purification.

[7]  Georges Mer,et al.  Structural Basis for the Methylation State-Specific Recognition of Histone H4-K20 by 53BP1 and Crb2 in DNA Repair , 2006, Cell.

[8]  Thomas A. Milne,et al.  A PHD finger of NURF couples histone H3 lysine 4 trimethylation with chromatin remodelling , 2006, Nature.

[9]  M. Stallcup,et al.  Developmentally Essential Protein Flightless I Is a Nuclear Receptor Coactivator with Actin Binding Activity , 2004, Molecular and Cellular Biology.

[10]  Randy J Read,et al.  Electronic Reprint Biological Crystallography Likelihood-enhanced Fast Rotation Functions Biological Crystallography Likelihood-enhanced Fast Rotation Functions , 2003 .

[11]  J. Northrop,et al.  Histone H3 Lysine 9 Methyltransferase G9a Is a Transcriptional Coactivator for Nuclear Receptors* , 2006, Journal of Biological Chemistry.

[12]  H. Kato,et al.  G9a histone methyltransferase plays a dominant role in euchromatic histone H3 lysine 9 methylation and is essential for early embryogenesis. , 2002, Genes & development.

[13]  Yang Shi,et al.  Recognition of unmethylated histone H3 lysine 4 links BHC80 to LSD1-mediated gene repression , 2007, Nature.

[14]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[15]  C. Allis,et al.  The language of covalent histone modifications , 2000, Nature.

[16]  A. Roopra,et al.  Localized domains of G9a-mediated histone methylation are required for silencing of neuronal genes. , 2004, Molecular cell.

[17]  K. Wright,et al.  PRDI-BF1 recruits the histone H3 methyltransferase G9a in transcriptional silencing , 2004, Nature Immunology.

[18]  Z. Otwinowski,et al.  Processing of X-ray diffraction data collected in oscillation mode. , 1997, Methods in enzymology.

[19]  E. Selker,et al.  In Vitro and in Vivo Analyses of a Phe/Tyr Switch Controlling Product Specificity of Histone Lysine Methyltransferases* , 2005, Journal of Biological Chemistry.

[20]  S. Grewal,et al.  Heterochromatin revisited , 2007, Nature Reviews Genetics.

[21]  C. Allis,et al.  Translating the Histone Code , 2001, Science.

[22]  Y. Shinkai,et al.  Zinc Finger Protein Wiz Links G9a/GLP Histone Methyltransferases to the Co-repressor Molecule CtBP* , 2006, Journal of Biological Chemistry.

[23]  Z. Duan,et al.  Gfi1 Coordinates Epigenetic Repression of p21Cip/WAF1 by Recruitment of Histone Lysine Methyltransferase G9a and Histone Deacetylase 1 , 2005, Molecular and Cellular Biology.

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

[25]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.