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PMID: 9806908 Published · ppublish English Journal Article

Cholesterol regulates oxysterol binding protein (OSBP) phosphorylation and Golgi localization in Chinese hamster ovary cells: correlation with stimulation of sphingomyelin synthesis by 25-hydroxycholesterol.

The Biochemical journal ·Vol. 336 ( Pt 1) ·1998-11-15 ·Pages 247-56

Storey MK, Byers DM, Cook HW, Ridgway ND

Abstract

Sphingomyelin (SM) and cholesterol content is positively correlated in cellular membranes, and in several pathological and experimental conditions there is evidence for coregulation. The potential role of oxysterols and oxysterol binding protein (OSBP) in mediating the coregulation of cholesterol and SM was examined using Chinese hamster ovary (CHO) and cholesterol auxotrophic, sterol regulatory defective (SRD) 6 cells. SRD 6 cells grown in the presence or absence of cholesterol for 24 h displayed a 30-50% reduction in SM synthesis compared with control CHO 7 cells. SM synthesis in CHO 7 and cholesterol-supplemented SRD 6 cells was stimulated 2-fold by 25-hydroxycholesterol, but cholesterol-starved SRD 6 cells were unresponsive. Basal and 25-hydroxycholesterol-stimulated SM synthesis was also inhibited in lovastatin-treated wild-type CHO-K1 cells. Lack of 25-hydroxycholesterol activation of SM synthesis in cholesterol-starved SRD 6 and lovastatin-treated CHO-K1 cells was correlated with dephosphorylation of OSBP. In SRD 6 cells, this was evident after 12 h of cholesterol depletion, it occurred equally at all phosphorylation sites and was exacerbated by 25-hydroxycholesterol. Unlike CHO 7 cells, where OSBP was observed in small vesicles and the cytoplasm, OSBP in cholesterol-starved SRD 6 cells was constitutively localized in the Golgi apparatus. Supplementation with non-lipoprotein cholesterol promoted redistribution to vesicles and the cytoplasm. Similarly, OSBP in CHO-K1 cells grown in delipidated serum was predominantly in the Golgi apparatus. Low-density lipoprotein (LDL) supplementation of CHO-K1 cells caused the redistribution of OSBP to the cytoplasm and small vesicles, and this effect was blocked by pharmacological agents ¿3-beta-[2-(diethylamino)ethoxy]androst-5-en-17-one and progesterone¿, which inhibited LDL cholesterol efflux from lysosomes. The results showed that localization of OSBP between the Golgi apparatus and a cytoplasmic/vesicular compartment was responsive to changes in cholesterol content and trafficking. In cholesterol depleted SRD 6 cells, this was accompanied by dephosphorylation of OSBP and attenuation of 25-hydroxycholesterol activation of SM synthesis.

MeSH Terms
Animals Anticholesteremic Agents/pharmacology CHO Cells Cholesterol/metabolism Cricetinae Cricetulus Fluorescent Antibody Technique, Indirect Golgi Apparatus/metabolism Hydroxycholesterols/pharmacology Lovastatin/pharmacology Phosphorylation Receptors, Steroid/metabolism Sphingomyelins/biosynthesis
Chemicals
Anticholesteremic Agents Hydroxycholesterols Receptors, Steroid Sphingomyelins oxysterol binding protein 25-hydroxycholesterol Cholesterol Lovastatin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Storey M K
Department of Pediatrics, Atlantic Research Centre, Dalhousie University, 5849 University Avenue, Halifax, Nova Scotia, Canada, B3H 4H7.
Byers D M
Cook H W
Ridgway N D
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3360247
Published
1998-11-15
Pages
247-56
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1219865
Subset
IM
External Links
PubMed source DOI: 10.1042/bj3360247 PMC Full Text View journal details
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