GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis

• Published in: Nature Vol. 459; no. 7245; pp. 455 - 459
• Main Authors: Fujiki, Ryoji, Chikanishi, Toshihiro, Hashiba, Waka, Ito, Hiroaki, Takada, Ichiro, Roeder, Robert G., Kitagawa, Hirochika, Kato, Shigeaki
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 21.05.2009; Nature Publishing Group
• Subjects: Acetylglucosamine - metabolism; Biological and medical sciences; Cell differentiation; Cell differentiation, maturation, development, hematopoiesis; Cell Lineage; Cell Nucleus - metabolism; Cell physiology; Cells; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Enzymes; Fundamental and applied biological sciences. Psychology; Genetic aspects; Granulocytes - cytology; Granulocytes - drug effects; Histone-Lysine N-Methyltransferase - chemistry; Histone-Lysine N-Methyltransferase - metabolism; Histones; HL-60 Cells; Humanities and Social Sciences; Humans; letter; Leukopoiesis - drug effects; Mass spectrometry; Methylation; Molecular and cellular biology; Monoclonal antibodies; multidisciplinary; Multiprotein Complexes - chemistry; Multiprotein Complexes - isolation & purification; Multiprotein Complexes - metabolism; N-Acetylglucosaminyltransferases - chemistry; N-Acetylglucosaminyltransferases - metabolism; Physiological aspects; Protein Structure, Tertiary; Proteins; Receptors, Retinoic Acid - metabolism; Retinoic Acid Receptor alpha; Science; Science (multidisciplinary); Threonine - metabolism; Tretinoin - pharmacology; United States; Posttranslational modification; Histone-lysine N-methyltransferase; Methyltransferases; Enzyme; Transferases; Retinoic acid; HL60 cell line; Granulopoiesis
• ISSN: 0028-0836; 1476-4687; 1476-4679
• DOI: 10.1038/nature07954

GlcNAcylation of chromatin proteins in granulopoiesis The SET-domain-containing MLL proteins methylate histone H3 on lysine 4 and contribute to gene activation. Here, MLL5 is shown to associate in a complex with the retinoic acid receptor RARα and to be modified by O -linked β- N -acetylglucosamine ( O -GlcNAc). Nuclear GlcNAcylation of MLL5 facilitates its activity as a co-activator of RARα target genes during granulopoiesis. This work points to an important role for GlcNAcylation in modifying chromatin proteins. The post-translational modifications of histone tails generate a 'histone code' that defines local and global chromatin states. Here it is shown that nuclear GlcNAcylation of a histone lysine methyltransferase, MLL5, by O -linked β- N -acetylglucosamine transferase, facilitates retinoic-acid-induced granulopoiesis. The post-translational modifications of histone tails generate a ‘histone code’ that defines local and global chromatin states 1 . The resultant regulation of gene function is thought to govern cell fate, proliferation and differentiation 2 . Reversible histone modifications such as methylation are under mutual controls to organize chromosomal events 3 , 4 . Among the histone modifications, methylation of specific lysine and arginine residues seems to be critical for chromatin configuration and control of gene expression 5 . Methylation of histone H3 lysine 4 (H3K4) changes chromatin into a transcriptionally active state 6 . Reversible modification of proteins by β- N -acetylglucosamine ( O -GlcNAc) in response to serum glucose levels regulates diverse cellular processes 7 , 8 , 9 . However, the epigenetic impact of protein GlcNAcylation is unknown. Here we report that nuclear GlcNAcylation of a histone lysine methyltransferase (HKMT), MLL5, by O -GlcNAc transferase facilitates retinoic-acid-induced granulopoiesis in human HL60 promyelocytes through methylation of H3K4. MLL5 is biochemically identified in a GlcNAcylation-dependent multi-subunit complex associating with nuclear retinoic acid receptor RARα (also known as RARA), serving as a mono- and di-methyl transferase to H3K4. GlcNAcylation at Thr 440 in the MLL5 SET domain evokes its H3K4 HKMT activity and co-activates RARα in target gene promoters. Increased nuclear GlcNAcylation by means of O -GlcNAc transferase potentiates retinoic-acid-induced HL60 granulopoiesis and restores the retinoic acid response in the retinoic-acid-resistant HL60-R2 cell line. Thus, nuclear MLL5 GlcNAcylation triggers cell lineage determination of HL60 through activation of its HKMT activity.