Home LiteratureArticle Details
PMID: 12370080 Published · ppublish English Journal Article

Inhibition of phosphatidylcholine synthesis induces expression of the endoplasmic reticulum stress and apoptosis-related protein CCAAT/enhancer-binding protein-homologous protein (CHOP/GADD153).

The Biochemical journal ·Vol. 369 ·No. Pt 3 ·2003-02-01 ·Pages 643-50

van der Sanden MH, Houweling M, van Golde LM, Vaandrager AB

Abstract

Inhibition of de novo synthesis of phosphatidylcholine (PC) by some anti-cancer drugs such as hexadecylphosphocholine leads to apoptosis in various cell lines. Likewise, in MT58, a mutant Chinese hamster ovary (CHO) cell line containing a thermo-sensitive mutation in CTP:phosphocholine cytidylyltransferase (CT), an important regulatory enzyme in the CDP-choline pathway, inhibition of PC synthesis causes PC depletion. Cellular perturbations like metabolic insults and unfolded proteins can be registered by the endoplasmic reticulum (ER) and result in ER stress responses, which can lead eventually to apoptosis. In this study we investigated the effect of PC depletion on the ER stress response and ER-related proteins. Shifting MT58 cells to the non-permissive temperature of 40 degrees C resulted in PC depletion via an inhibition of CT within 24 h. Early apoptotic features appeared in several cells around 30 h, and most cells were apoptotic within 48 h. The temperature shift in MT58 led to an increase of pro-apoptotic CCAAT/enhancer-binding protein-homologous protein (CHOP; also known as GADD153) after 16 h, to a maximum at 24 h. Incubation of wild-type CHO-K1 or CT-expressing MT58 cells at 40 degrees C did not induce differences in CHOP protein levels in time. In contrast, expression of the ER chaperone BiP/GRP78, induced by an increase in misfolded/unfolded proteins, and caspase 12, a protease specifically involved in apoptosis that results from stress in the ER, did not differ between MT58 and CHO-K1 cells in time when cultured at 40 degrees C. Furthermore, heat-shock protein 70, a protein that is stimulated by accumulation of abnormal proteins and heat stress, displayed similar expression patterns in MT58 and K1 cells. These results suggest that PC depletion in MT58 induces the ER-stress-related protein CHOP, without raising a general ER stress response.

MeSH Terms
Animals Apoptosis/physiology CCAAT-Enhancer-Binding Proteins/metabolism CHO Cells Carrier Proteins/metabolism Caspase 12 Caspases/metabolism Choline-Phosphate Cytidylyltransferase/genetics,metabolism Cricetinae Endoplasmic Reticulum/metabolism Endoplasmic Reticulum Chaperone BiP Heat-Shock Proteins Molecular Chaperones/metabolism Mutation Phosphatidylcholines/biosynthesis Stress, Physiological Temperature Transcription Factor CHOP Transcription Factors/metabolism
Chemicals
CCAAT-Enhancer-Binding Proteins Carrier Proteins Endoplasmic Reticulum Chaperone BiP Heat-Shock Proteins Molecular Chaperones Phosphatidylcholines Transcription Factors Transcription Factor CHOP Choline-Phosphate Cytidylyltransferase Caspase 12 Caspases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
van der Sanden Michiel H M
Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine and Institute of Biomembranes, University of Utrecht, The Netherlands. [email protected]
Houweling Martin
van Golde Lambert M G
Vaandrager Arie B
References (57)
57 references, click to expand
  1. Activation of caspase-12, an endoplastic reticulum (ER) resident caspase, through tumor necrosis factor receptor-associated factor 2-dependent mechanism in response to the ER stress.
    J Biol Chem. 2001 Apr 27;276(17):13935-40 PMID: 11278723
  2. Eukaryotic phospholipid biosynthesis.
    Annu Rev Biochem. 1995;64:315-43 PMID: 7574485
  3. Autoradiographic detection of animal cell membrane mutants altered in phosphatidylcholine synthesis.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5192-6 PMID: 6254065
  4. Thermolabile CDP-choline synthetase in an animal cell mutant defective in lecithin formation.
    J Biol Chem. 1981 Jul 25;256(14):7388-93 PMID: 6265447
  5. Stress signaling from the lumen of the endoplasmic reticulum: coordination of gene transcriptional and translational controls.
    Genes Dev. 1999 May 15;13(10):1211-33 PMID: 10346810
  6. Induction of ceramide-mediated apoptosis by the anticancer phospholipid analog, hexadecylphosphocholine.
    J Biol Chem. 1998 May 1;273(18):11025-31 PMID: 9556584
  7. Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-beta.
    Nature. 2000 Jan 6;403(6765):98-103 PMID: 10638761
  8. GADD153/CHOP, a DNA damage-inducible protein, reduced CAAT/enhancer binding protein activities and increased apoptosis in 32D c13 myeloid cells.
    Cancer Res. 1996 Jul 15;56(14):3250-6 PMID: 8764117
  9. Amino acids control mammalian gene transcription: activating transcription factor 2 is essential for the amino acid responsiveness of the CHOP promoter.
    Mol Cell Biol. 2000 Oct;20(19):7192-204 PMID: 10982836
  10. Interactions of misfolded influenza virus hemagglutinin with binding protein (BiP).
    J Cell Biol. 1989 Jun;108(6):2117-26 PMID: 2738090
  11. Stress-induced binding of the transcriptional factor CHOP to a novel DNA control element.
    Mol Cell Biol. 1996 Apr;16(4):1479-89 PMID: 8657121
  12. Expression of wild-type and mutant rat liver CTP: phosphocholine cytidylyltransferase in a cytidylyltransferase-deficient Chinese hamster ovary cell line.
    Arch Biochem Biophys. 1994 May 15;311(1):107-16 PMID: 8185307
  13. Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase.
    Nature. 1999 Jan 21;397(6716):271-4 PMID: 9930704
  14. Hsp70 exerts its anti-apoptotic function downstream of caspase-3-like proteases.
    EMBO J. 1998 Nov 2;17(21):6124-34 PMID: 9799222
  15. Glutamine deprivation induces the expression of GADD45 and GADD153 primarily by mRNA stabilization.
    J Biol Chem. 1999 Oct 1;274(40):28645-51 PMID: 10497233
  16. Inhibition of phosphatidylcholine biosynthesis following induction of apoptosis in HL-60 cells.
    J Biol Chem. 1999 Jul 9;274(28):19686-92 PMID: 10391908
  17. Fas- or ceramide-induced apoptosis is mediated by a Rac1-regulated activation of Jun N-terminal kinase/p38 kinases and GADD153.
    J Biol Chem. 1997 Aug 29;272(35):22173-81 PMID: 9268362
  18. CHOP is implicated in programmed cell death in response to impaired function of the endoplasmic reticulum.
    Genes Dev. 1998 Apr 1;12(7):982-95 PMID: 9531536
  19. Stress-induced phosphorylation and activation of the transcription factor CHOP (GADD153) by p38 MAP Kinase.
    Science. 1996 May 31;272(5266):1347-9 PMID: 8650547
  20. Regulation of CTP:phosphocholine cytidylyltransferase by amphitropism and relocalization.
    Trends Biochem Sci. 2000 Sep;25(9):441-7 PMID: 10973058
  21. Amino acid limitation regulates CHOP expression through a specific pathway independent of the unfolded protein response.
    FEBS Lett. 1999 Apr 9;448(2-3):211-6 PMID: 10218478
  22. CHOP gene expression in response to endoplasmic-reticular stress requires NFY interaction with different domains of a conserved DNA-binding element.
    Nucleic Acids Res. 2000 Dec 15;28(24):4987-97 PMID: 11121490
  23. Tripartite management of unfolded proteins in the endoplasmic reticulum.
    Cell. 2000 May 26;101(5):451-4 PMID: 10850487
  24. Coupling endoplasmic reticulum stress to the cell death program. Mechanism of caspase activation.
    J Biol Chem. 2001 Sep 7;276(36):33869-74 PMID: 11448953
  25. Quantitative analysis of phospholipids by thin-layer chromatography and phosphorus analysis of spots.
    Lipids. 1966 Jan;1(1):85-6 PMID: 17805690
  26. Coordination of membrane phospholipid synthesis with the cell cycle.
    J Biol Chem. 1994 Feb 4;269(5):3858-67 PMID: 8106431
  27. Distribution of CTP:phosphocholine cytidylyltransferase (CCT) isoforms. Identification of a new CCTbeta splice variant.
    J Biol Chem. 1999 Sep 17;274(38):26992-7001 PMID: 10480912
  28. The antiproliferative effect of hexadecylphosphocholine toward HL60 cells is prevented by exogenous lysophosphatidylcholine.
    Biochim Biophys Acta. 1998 Jan 5;1389(1):1-12 PMID: 9443598
  29. Cell cycle regulation of membrane phospholipid metabolism.
    J Biol Chem. 1996 Aug 23;271(34):20219-22 PMID: 8702749
  30. Expression of phosphatidylethanolamine N-methyltransferase-2 cannot compensate for an impaired CDP-choline pathway in mutant Chinese hamster ovary cells.
    J Biol Chem. 1995 Jul 7;270(27):16277-82 PMID: 7608195
  31. Role of the human heat shock protein hsp70 in protection against stress-induced apoptosis.
    Mol Cell Biol. 1997 Sep;17(9):5317-27 PMID: 9271409
  32. Mammalian stress response: induction of the glucose-regulated protein family.
    Curr Opin Cell Biol. 1992 Apr;4(2):267-73 PMID: 1599691
  33. Heat-shock protein 70 antisense oligomers enhance proteasome inhibitor-induced apoptosis.
    Biochem J. 1999 Dec 1;344 Pt 2:477-85 PMID: 10567231
  34. Ceramide: an intracellular signal for apoptosis.
    Trends Biochem Sci. 1995 Feb;20(2):73-7 PMID: 7701566
  35. Effect of amino acid limitation on the expression of 19 genes in rat hepatoma cells.
    FASEB J. 1994 May;8(8):538-44 PMID: 8181673
  36. How cytidylyltransferase uses an amphipathic helix to sense membrane phospholipid composition.
    Biochem Soc Trans. 1998 Aug;26(3):539-44 PMID: 9765910
  37. The presence of malfolded proteins in the endoplasmic reticulum signals the induction of glucose-regulated proteins.
    Nature. 1988 Mar 31;332(6163):462-4 PMID: 3352747
  38. Brefeldin A, thapsigargin, and AIF4- stimulate the accumulation of GRP78 mRNA in a cycloheximide dependent manner, whilst induction by hypoxia is independent of protein synthesis.
    J Cell Physiol. 1992 Sep;152(3):545-52 PMID: 1506413
  39. Animal cells dependent on exogenous phosphatidylcholine for membrane biogenesis.
    Proc Natl Acad Sci U S A. 1982 Mar;79(6):1698-702 PMID: 6281780
  40. Phosphatidylcholine breakdown and signal transduction.
    Biochim Biophys Acta. 1994 Apr 14;1212(1):26-42 PMID: 8155724
  41. The molecular response to reductive stress in LLC-PK1 renal epithelial cells: coordinate transcriptional regulation of gadd153 and grp78 genes by thiols.
    Cell Stress Chaperones. 1997 Mar;2(1):31-40 PMID: 9250393
  42. A rapid method of total lipid extraction and purification.
    Can J Biochem Physiol. 1959 Aug;37(8):911-7 PMID: 13671378
  43. Gadd45 and Gadd153 messenger RNA levels are increased during hypoxia and after exposure of cells to agents which elevate the levels of the glucose-regulated proteins.
    Cancer Res. 1992 Jul 1;52(13):3814-7 PMID: 1617653
  44. Distinct roles of activating transcription factor 6 (ATF6) and double-stranded RNA-activated protein kinase-like endoplasmic reticulum kinase (PERK) in transcription during the mammalian unfolded protein response.
    Biochem J. 2002 Sep 1;366(Pt 2):585-94 PMID: 12014989
  45. Calcium ionophore A23187 induces expression of the growth arrest and DNA damage inducible CCAAT/enhancer-binding protein (C/EBP)-related gene, gadd153. Ca2+ increases transcriptional activity and mRNA stability.
    J Biol Chem. 1992 Oct 5;267(28):20465-70 PMID: 1400365
  46. Sphingomyelin synthase and PKC activation.
    Science. 1989 Nov 24;246(4933):1050 PMID: 2555921
  47. Apoptosis triggered by 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine is prevented by increased expression of CTP:phosphocholine cytidylyltransferase.
    J Biol Chem. 1998 Jan 23;273(4):2169-73 PMID: 9442058
  48. CTP:phosphocholine cytidylyltransferase.
    Biochim Biophys Acta. 1997 Sep 4;1348(1-2):79-90 PMID: 9370319
  49. Structure of the membrane binding domain of CTP:phosphocholine cytidylyltransferase.
    Biochemistry. 1996 Sep 17;35(37):11975-84 PMID: 8810902
  50. Identification of novel stress-induced genes downstream of chop.
    EMBO J. 1998 Jul 1;17(13):3619-30 PMID: 9649432
  51. Phosphatidylcholine cycle and regulation of phosphatidylcholine biosynthesis by enzyme translocation.
    Biochim Biophys Acta. 1994 May 13;1212(2):137-51 PMID: 8180240
  52. Competitive inhibition of choline phosphotransferase by geranylgeraniol and farnesol inhibits phosphatidylcholine synthesis and induces apoptosis in human lung adenocarcinoma A549 cells.
    J Biol Chem. 1998 Oct 2;273(40):26179-86 PMID: 9748300
  53. Regulation of phosphatidylcholine biosynthesis.
    Prog Lipid Res. 1990;29(2):87-105 PMID: 1965552
  54. CHOP, a novel developmentally regulated nuclear protein that dimerizes with transcription factors C/EBP and LAP and functions as a dominant-negative inhibitor of gene transcription.
    Genes Dev. 1992 Mar;6(3):439-53 PMID: 1547942
  55. Signals from the stressed endoplasmic reticulum induce C/EBP-homologous protein (CHOP/GADD153).
    Mol Cell Biol. 1996 Aug;16(8):4273-80 PMID: 8754828
  56. Colony-stimulating factor 1 regulates CTP: phosphocholine cytidylyltransferase mRNA levels.
    J Biol Chem. 1991 Sep 5;266(25):16261-4 PMID: 1653227
  57. A genetic defect in phosphatidylcholine biosynthesis triggers apoptosis in Chinese hamster ovary cells.
    J Biol Chem. 1996 Jun 21;271(25):14668-71 PMID: 8663247
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/BJ20020285
Email
[email protected]
Published
2003-02-01
Pages
643-50
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1223098
Subset
IM
External Links
PubMed source DOI: 10.1042/BJ20020285 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2