Regulation of phosphatidylcholine homeostasis by calcium-independent phospholipase A2.
Phosphatidylcholine (PtdCho) is the most abundant phospholipid in mammalian cell membranes and is essential for cell viability. The levels of this lipid must be tightly controlled to maintain homeostasis. Therefore, changes in the rate of PtdCho synthesis are generally balanced by changes in PtdCho catabolism and vice versa. It is commonly accepted that the rate of PtdCho synthesis is regulated by CTP:phosphocholine cytidylyltransferase (CT). However, it is not certain if PtdCho mass is regulated by specific catabolic enzyme(s). Our goal is to determine if PtdCho homeostasis is regulated by a phospholipase A2 (PLA2). To this end, we have prepared Chinese hamster ovary (CHO) cell lines that overexpress CT. CT activity is 7-10-fold higher in the transfected cells than in parental CHO cells. This increase in CT activity is associated with increases in both PtdCho synthesis and PtdCho catabolism. Glycerophosphocholine is the PtdCho catabolite that accumulates in the transfected cells, which suggests that PtdCho turnover is mediated by a phospholipase A2 (PLA2). Indeed, higher levels of calcium-independent PLA2 activity are measured in the cytosols of the CHO cells that overexpress CT, compared to parental CHO cells. The elevated calcium-independent PLA2 activity is associated with increases in the expression of the 80-kDa calcium-independent PLA2 (iPLA2). Together, these data suggest that the 80-kDa iPLA2 may be modulated in response to changes in PtdCho levels and therefore is involved in the regulation of PtdCho homeostasis in CHO cells.
Barbour
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SE
Department of Microbiology and Immunology, Virginia Commonwealth University, Box 980678, Richmond, VA 23298-0678, USA. sbarbour@hsc.vcu.edu
Kapur
A
A
Deal
C L
CL
D000818:Animals; D016466:CHO Cells; D002470:Cell Survival; D019916:Choline-Phosphate Cytidylyltransferase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D015870:Gene Expression; D005997:Glycerylphosphorylcholine / Q000378:metabolism; D054522:Group VI Phospholipases A2; D006706:Homeostasis; D015151:Immunoblotting; D010713:Phosphatidylcholines / Q000096:biosynthesis / Q000378:metabolism*; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D014162:Transfection
D010713:Phosphatidylcholines; D005997:Glycerylphosphorylcholine; D019916:Choline-Phosphate Cytidylyltransferase; D010741:Phospholipases A; D054522:Group VI Phospholipases A2; D054467:Phospholipases A2