Lipoprotein deprivation stimulates transcription of the CTP:phosphocholine cytidylyltransferase gene.
We examined the effect of lipoprotein deprivation on the expression of the rate-regulatory enzyme involved in phosphatidylcholine (PtdCho) synthesis, phosphocholine cytidylyltransferase (CCT), within an alveolar type II epithelial cell line (MLE-12). Compared with cells exposed to 10% fetal bovine serum (FBS, control), cells cultured with lipoprotein-deficient serum (LPDS) for 72 h had a 150% increase in CCT activity. Stimulation of CCT activity after LPDS exposure was associated with a 2-fold increase in immunoreactive CCT content and a corresponding increase in [(35)S]methionine incorporation into newly synthesized CCT. LPDS induction of CCT protein was reversible, as it was suppressed to baseline levels by the addition of low density lipoproteins to the culture medium. Northern blotting revealed that LPDS increased CCT mRNA levels 2-fold compared with control. The induction of CCT mRNA by LPDS was not associated with an increase in mRNA half-life. Nuclear run-on assays revealed that LPDS-induced expression of CCT was due, at least in part, to an increase in gene transcription. These studies reveal that lipoprotein deprivation upregulates the activity of a key enzyme involved in the PtdCho biosynthetic pathway. LPDS induction of CCT protein might serve as a novel compensatory mechanism in response to lipid deprivation by increasing cellular transcription of the CCT gene.
Ryan
A J
AJ
Department of Internal Medicine and Veterans Affairs Medical Center, University of Iowa College of Medicine, Iowa City, IA 52242, USA.
McCoy
D M
DM
Mathur
S N
SN
Field
F J
FJ
Mallampalli
R K
RK
D000818:Animals; D001769:Blood; D015152:Blotting, Northern; D019916:Choline-Phosphate Cytidylyltransferase / Q000096:biosynthesis / Q000235:genetics*; D003470:Culture Media; D004847:Epithelial Cells / Q000201:enzymology; D006207:Half-Life; D015151:Immunoblotting; D008074:Lipoproteins / Q000097:blood / Q000172:deficiency*; D010713:Phosphatidylcholines / Q000096:biosynthesis; D011650:Pulmonary Alveoli / Q000201:enzymology; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D014158:Transcription, Genetic
D003470:Culture Media; D008074:Lipoproteins; D010713:Phosphatidylcholines; D012333:RNA, Messenger; D019916:Choline-Phosphate Cytidylyltransferase