![]() |
|
|
Vol. 19, Issue 10, 4130-4140, October 2008
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||


*Department of Biological Chemistry, The Johns Hopkins School of Medicine, Baltimore, MD 21205;
Momenta Pharmaceuticals, Cambridge, MA 02142; and
Department of Chemistry and Chemical Biology, Rutgers, The State University of New Jersey, Piscataway, NJ 08554
Submitted November 15, 2007;
Revised July 9, 2008;
Accepted July 10, 2008
Monitoring Editor: M. Bishr Omary
O-linked β-N-acetylglucosamine (O-GlcNAc) is a highly dynamic intracellular protein modification responsive to stress, hormones, nutrients, and cell cycle stage. Alterations in O-GlcNAc addition or removal (cycling) impair cell cycle progression and cytokinesis, but the mechanisms are not well understood. Here, we demonstrate that the enzymes responsible for O-GlcNAc cycling, O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA) are in a transient complex at M phase with the mitotic kinase Aurora B and protein phosphatase 1. OGT colocalized to the midbody during telophase with Aurora B. Furthermore, these proteins coprecipitated with each other in a late mitotic extract. The complex was stable under Aurora inhibition; however, the total cellular levels of O-GlcNAc were increased and the localization of OGT was decreased at the midbody after Aurora inhibition. Vimentin, an intermediate filament protein, is an M phase substrate for both Aurora B and OGT. Overexpression of OGT or OGA led to defects in mitotic phosphorylation on multiple sites, whereas OGT overexpression increased mitotic GlcNAcylation of vimentin. OGA inhibition caused a decrease in vimentin late mitotic phosphorylation but increased GlcNAcylation. Together, these data demonstrate that the O-GlcNAc cycling enzymes associate with kinases and phosphatases at M phase to regulate the posttranslational status of vimentin.
Address correspondence to: Gerald W. Hart (gwhart{at}jhmi.edu)
Abbreviations used: GFP, green fluorescent protein; GT, O-GlcNAc-thiazoline; OGA, O-GlcNAcase; O-GlcNAc, O-linked β-N-acetylglucosamine; OGT, O-GlcNAc transferase; PP1, protein phosphatase 1; ZM, ZM447439.
Related articles in Mol. Biol. Cell:
This article has been cited by other articles:
![]() |
B. Laczy, B. G. Hill, K. Wang, A. J. Paterson, C. R. White, D. Xing, Y.-F. Chen, V. Darley-Usmar, S. Oparil, and J. C. Chatham Protein O-GlcNAcylation: a new signaling paradigm for the cardiovascular system Am J Physiol Heart Circ Physiol, January 1, 2009; 296(1): H13 - H28. [Abstract] [Full Text] [PDF] |
||||