コンテンツへスキップ
Merck

Undetectable histone O-GlcNAcylation in mammalian cells.

Epigenetics (2015-06-16)
Jessica Gagnon, Salima Daou, Natalia Zamorano, Nicholas V G Iannantuono, Ian Hammond-Martel, Nazar Mashtalir, Eric Bonneil, Hugo Wurtele, Pierre Thibault, El Bachir Affar
要旨

O-GlcNAcylation is a posttranslational modification catalyzed by the O-Linked N-acetylglucosamine (O-GlcNAc) transferase (OGT) and reversed by O-GlcNAcase (OGA). Numerous transcriptional regulators, including chromatin modifying enzymes, transcription factors, and co-factors, are targeted by O-GlcNAcylation, indicating that this modification is central for chromatin-associated processes. Recently, OGT-mediated O-GlcNAcylation was reported to be a novel histone modification, suggesting a potential role in directly coordinating chromatin structure and function. In contrast, using multiple biochemical approaches, we report here that histone O-GlcNAcylation is undetectable in mammalian cells. Conversely, O-GlcNAcylation of the transcription regulators Host Cell Factor-1 (HCF-1) and Ten-Eleven Translocation protein 2 (TET2) could be readily observed. Our study raises questions on the occurrence and abundance of O-GlcNAcylation as a histone modification in mammalian cells and reveals technical complications regarding the detection of genuine protein O-GlcNAcylation. Therefore, the identification of the specific contexts in which histone O-GlcNAcylation might occur is still to be established.

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
モノクローナル抗FLAG® M2抗体 マウス宿主抗体, clone M2, purified immunoglobulin (Purified IgG1 subclass), buffered aqueous solution (10 mM sodium phosphate, 150 mM NaCl, pH 7.4, containing 0.02% sodium azide)
Sigma-Aldrich
抗アクチン抗体、クローンC4, ascites fluid, clone C4, Chemicon®
Sigma-Aldrich
抗グルタチオン-S-トランスフェラーゼ (GST)-アガロース ウサギ宿主抗体, IgG fraction of antiserum, PBS suspension