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Microinjection of recombinant O-GlcNAc transferase potentiates Xenopus oocytes M-phase entry
Authors:Dehennaut Vanessa  Hanoulle Xavier  Bodart Jean-François  Vilain Jean-Pierre  Michalski Jean-Claude  Landrieu Isabelle  Lippens Guy  Lefebvre Tony
Affiliation:a UMR-CNRS 8576, UGSF, USTL, IFR 147, 59655 Villeneuve d’Ascq, France
b EA 4020, Laboratoire de Régulation des Signaux de Division, USTL, IFR147, Villeneuve d’Ascq, France
Abstract:In order to understand the importance of the cytosolic and nuclear-specific O-linked N-acetylglucosaminylation (O-GlcNAc) on cell cycle regulation, we recently reported that inhibition of O-GlcNAc transferase (OGT) delayed or blocked Xenopus laevis oocyte germinal vesicle breakdown (GVBD). Here, we show that increased levels of the long OGT isoform (ncOGT) accelerate X. laevis oocyte GVBD. A N-terminally truncated isoform (sOGT) with a similar in vitro catalytic activity towards a synthetic CKII-derived peptide had no effect, illustrating the important role played by the N-terminal tetratrico-peptide repeats. ncOGT microinjection in the oocytes increases both the speed and extent of O-GlcNAc addition, leads to a quicker activation of the MPF and MAPK pathways and finally results in a faster GVBD. Microinjection of anti-OGT antibodies leads to a delay of the GVBD kinetics. Our results hence demonstrate that OGT is a key molecule for the timely progression of the cell cycle.
Keywords:OGT   O-GlcNAc   Xenopus laevis   Oocyte   Cell cycle
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