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J Biol Chem, Vol. 274, Issue 37, 26240-26248, September 10, 1999
G1 Transition
From the Institute of Molecular Biology and Biochemistry and the
Biochemistry Program, Simon Fraser University, Burnaby,
British Columbia V5A 1S6, Canada
The transition from quiescence
(G0) into the cell division cycle is marked by
accelerated phospholipid turnover. We examined the rates of
phosphatidylcholine (PC) synthesis and the activity, membrane affinity,
and intracellular localization of the rate-limiting enzyme in the
synthesis of PC, CTP:phosphocholine cytidylyltransferase (CT) during
this transition. The addition of serum to quiescent IIC9 fibroblasts
resulted in a wave of PC synthesis beginning at ~10 min, peaking at
~3 h with a >10-fold increase in rate, and declining to near basal
rates by 10 h. CT activity, monitored in situ, was
elevated ~3-fold between 1 and 2 h postserum. Neither CT mass
nor its phosphorylation state changed during the surge in PC synthesis
and CT activity. On the other hand, the ratio of particulate/soluble CT
surged and then receded in concert with the wave of PC synthesis.
During quiescence, CT was confined to the nucleus, as assessed by
indirect immunofluorescence. Within 10 min after serum stimulation, a
portion of the CT fluorescence appeared in the cytoplasm, where it
intensified until ~4 h postserum. Thereafter, the cytoplasmic CT
signal waned, while the nuclear signal increased, and by 8 h CT
was once again predominantly nuclear. The dynamics of CT's apparent
translocation in and out of the nucleus paralleled the wave of PC
synthesis and the solubility changes of CT. Cytoplasmic CT co-localized
with BiP, a resident endoplasmic reticulum protein, in a double
labeling experiment. These data suggest that the wave of PC synthesis
that accompanies the G0 
G1 transition is
regulated by the coordinated changes in CT activity, membrane affinity,
and intracellular distribution. We describe for the first time a
redistribution of CT from the nucleus to the ER that correlates with an
activation of the enzyme. We propose that this movement is required for
the stimulation of PC synthesis during entry into the cell cycle.
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