The site-specific installation of methyl-lysine analogs into recombinant histones

被引:378
作者
Simon, Matthew D.
Chu, Feixia
Racki, Lisa R.
de la Cruz, Cecile C.
Burlingame, Alma L.
Panning, Barbara
Narlikar, Geeta J.
Shokat, Kevan M. [1 ]
机构
[1] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA
[2] Univ Calif San Francisco, Dept Biochem, San Francisco, CA 94143 USA
[3] Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA
[4] Univ Calif San Francisco, Howard Hughes Med Inst, Dept Cellular & Mol Pharmacol, San Francisco, CA 94143 USA
关键词
D O I
10.1016/j.cell.2006.12.041
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Histone lysine residues can be mono-, di-, or trimethylated. These posttranslational modifications regulate the affinity of effector proteins and may also impact chromatin structure independent of their role as adaptors. In order to study histone lysine methylation, particularly in the context of chromatin, we have developed a chemical approach to install analogs of methyl lysine into recombinant proteins. This approach allows for the rapid generation of large quantities of histones in which the site and degree of methylation can be specified. We demonstrate that these methyl-lysine analogs (MLAs) are functionally similar to their natural counterparts. These methylated histones, were used to examine the influence of specific lysine methylation on the binding of effecter proteins and the rates of nucleosme remodeling. This simple method of introducing site-specific and degree-specific methylation into recombinant histones provides a powerful tool to investigate the biochemical mechanisms by which lysine methylation influences chromatin structure and function.
引用
收藏
页码:1003 / 1012
页数:10
相关论文
共 45 条
[11]   Facile synthesis of site-specifically acetylated and methylated histone proteins: Reagents for evaluation of the histone code hypothesis [J].
He, S ;
Bauman, D ;
Davis, JS ;
Loyola, A ;
Nishioka, K ;
Gronlund, JL ;
Reinberg, D ;
Meng, FY ;
Kelleher, N ;
McCafferty, DG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2003, 100 (21) :12033-12038
[12]   Human ACF1 alters the remodeling strategy of SNF2h [J].
He, Xi ;
Fan, Hua-Ying ;
Narlikar, Geeta J. ;
Kingston, Robert E. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2006, 281 (39) :28636-28647
[13]   Aminoethylation in model peptides reveals conditions for maximizing thiol specificity [J].
Hopkins, CE ;
Hernandez, G ;
Lee, JP ;
Tolan, DR .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2005, 443 (1-2) :1-10
[14]  
ITANO HA, 1972, J BIOL CHEM, V247, P4819
[15]   The chromo and SET domains of the Clr4 protein are essential for silencing in fission yeast [J].
Ivanova, AV ;
Bonaduce, MJ ;
Ivanov, SV ;
Klar, AJS .
NATURE GENETICS, 1998, 19 (02) :192-195
[16]   Structure of HP1 chromodomain bound to a lysine 9-methylated histone H3 tail [J].
Jacobs, SA ;
Khorasanizadeh, S .
SCIENCE, 2002, 295 (5562) :2080-2083
[17]  
Kenyon G L, 1977, Methods Enzymol, V47, P407
[18]   An epigenetic road map for histone lysine methylation [J].
Lachner, M ;
O'Sullivan, RJ ;
Jenuwein, T .
JOURNAL OF CELL SCIENCE, 2003, 116 (11) :2117-2124
[19]   Role of protein methylation in regulation of transcription [J].
Lee, DY ;
Teyssier, C ;
Strahl, BD ;
Stallcup, MR .
ENDOCRINE REVIEWS, 2005, 26 (02) :147-170
[20]   Molecular basis for site-specific read-out of histone H3K4me3 by the BPTF PHD finger of NURF [J].
Li, Haitao ;
Ilin, Serge ;
Wang, Wooikoon ;
Duncan, Elizabeth M. ;
Wysocka, Joanna ;
Allis, C. David ;
Patel, Dinshaw J. .
NATURE, 2006, 442 (7098) :91-95