Structural and functional analysis of SET8, a histone H4 Lys-20 methyltransferase.
SET8 (also known as PR-SET7) is a histone H4-Lys-20-specific methyltransferase that is implicated in cell-cycle-dependent transcriptional silencing and mitotic regulation in metazoans. Herein we report the crystal structure of human SET8 (hSET8) bound to a histone H4 peptide bearing Lys-20 and the product cofactor S-adenosylhomocysteine. Histone H4 intercalates in the substrate-binding cleft as an extended parallel beta-strand. Residues preceding Lys-20 in H4 engage in an extensive array of salt bridge, hydrogen bond, and van der Waals interactions with hSET8, while the C-terminal residues bind through predominantly hydrophobic interactions. Mutational analysis of both the substrate-binding cleft and histone H4 reveals that interactions with residues in the N and C termini of the H4 peptide are critical for conferring substrate specificity. Finally, analysis of the product specificity indicates that hSET8 is a monomethylase, consistent with its role in the maintenance of Lys-20 monomethylation during cell division.
Pubmed ID: 15933070 RIS Download
Binding Sites | Catalytic Domain | Crystallography, X-Ray | Histone-Lysine N-Methyltransferase | Histones | Humans | In Vitro Techniques | Models, Molecular | Mutagenesis, Site-Directed | Protein Conformation | Recombinant Proteins | S-Adenosylhomocysteine | Static Electricity | Substrate Specificity