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Molecules 2018, 23(6), 1275; https://doi.org/10.3390/molecules23061275

O-Linked N-Acetylglucosamine Transiently Elevates in HeLa Cells during Mitosis

1
Department of Laboratory Medicine, Medical School, University of Pécs, Pécs H7624, Hungary
2
Department of Biophysics, Medical School, University of Pécs, Pécs H7624, Hungary
3
Institute of Pharmacology and Toxicology, Paracelsus Medical University, Salzburg 5020, Austria
4
János Szentágothai Research Centre, University of Pécs, Pécs H7624, Hungary
*
Author to whom correspondence should be addressed.
Academic Editors: Franz-Georg Hanisch and Isabelle Breloy
Received: 27 April 2018 / Revised: 19 May 2018 / Accepted: 24 May 2018 / Published: 26 May 2018
(This article belongs to the Special Issue Functional Roles of Protein O-Glycosylation)
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Abstract

O-linked N-acetylglucosamine (O-GlcNAc) is a dynamic post-translational modification of serine and threonine residues on nuclear and cytoplasmic proteins. O-GlcNAc modification influences many cellular mechanisms, including carbohydrate metabolism, signal transduction and protein degradation. Multiple studies also showed that cell cycle might be modulated by O-GlcNAc. Although the role of O-GlcNAc in the regulation of some cell cycle processes such as mitotic spindle organization or histone phosphorylation is well established, the general behaviour of O-GlcNAc regulation during cell cycle is still controversial. In this study, we analysed the dynamic changes of overall O-GlcNAc levels in HeLa cells using double thymidine block. O-GlcNAc levels in G1, S, G2 and M phase were measured. We observed that O-GlcNAc levels are significantly increased during mitosis in comparison to the other cell cycle phases. However, this change could only be detected when mitotic cells were enriched by harvesting round shaped cells from the G2/M fraction of the synchronized cells. Our data verify that O-GlcNAc is elevated during mitosis, but also emphasize that O-GlcNAc levels can significantly change in a short period of time. Thus, selection and collection of cells at specific cell-cycle checkpoints is a challenging, but necessary requirement for O-GlcNAc studies. View Full-Text
Keywords: cell cycle; cell synchronization; mitosis; O-GlcNAc; post-translational modifications cell cycle; cell synchronization; mitosis; O-GlcNAc; post-translational modifications
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Fisi, V.; Kátai, E.; Orbán, J.; Dossena, S.; Miseta, A.; Nagy, T. O-Linked N-Acetylglucosamine Transiently Elevates in HeLa Cells during Mitosis. Molecules 2018, 23, 1275.

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