Microinjection of recombinant O-GlcNAc transferase potentiates Xenopus oocytes M-phase entry

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...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2008-05, Vol.369 (2), p.539-546
Main Authors: Dehennaut, Vanessa, Hanoulle, Xavier, Bodart, Jean-François, Vilain, Jean-Pierre, Michalski, Jean-Claude, Landrieu, Isabelle, Lippens, Guy, Lefebvre, Tony
Format: Article
Language:English
Subjects:
60 APPLIED LIFE SCIENCES
Acetylglucosamine - metabolism
Animals
ANTIBODIES
CELL CYCLE
Cell Division - drug effects
Cell Division - physiology
Cells, Cultured
GENE REGULATION
IN VITRO
INHIBITION
Life Sciences
Microinjections
MOLECULES
N-Acetylglucosaminyltransferases - administration & dosage
N-Acetylglucosaminyltransferases - genetics
N-Acetylglucosaminyltransferases - metabolism
O-GlcNAc
OGT
Oocyte
OOCYTES
Oocytes - cytology
Oocytes - drug effects
Oocytes - enzymology
PEPTIDES
Recombinant Proteins - administration & dosage
Xenopus laevis
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Summary: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.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2008.02.063