© The Rockefeller University Press,
0021-9525/1997//1411 $5.00
The Journal of Cell Biology, Volume 139, Number 6,
, 1997 1411-1418
Ceramide Accumulation Uncovers a Cycling Pathway for the cis-Golgi Network Marker, Infectious Bronchitis Virus M Protein
Michael Maceyka and
Carolyn E. Machamer
Department of Cell Biology and Anatomy, The Johns Hopkins School of Medicine, Baltimore, Maryland 21205
The M glycoprotein from the avian coronavirus, infectious bronchitis virus (IBV), contains information for localization to the cis-Golgi network in its first transmembrane domain. We hypothesize that localization to the Golgi complex may depend in part on specific interactions between protein transmembrane domains and membrane lipids. Because the site of sphingolipid synthesis overlaps the localization of IBV M, we asked whether perturbation of sphingolipids affected localization of IBV M. Short-term treatment with two inhibitors of sphingolipid synthesis had no effect on localization of IBV M or other Golgi markers. Thus, ongoing synthesis of these lipids was not required for proper localization. Surprisingly, a third inhibitor, d,l-threo-1-phenyl-2-decanoylamino-3-morpholino- 1-propanol (PDMP), shifted the steady-state distribution of IBV M from the Golgi complex to the ER. This effect was rapid and reversible and was also observed for ERGIC-53 but not for Golgi stack proteins. At the concentration of PDMP used, conversion of ceramide into both glucosylceramide and sphingomyelin was inhibited. Pretreatment with upstream inhibitors partially reversed the effects of PDMP, suggesting that ceramide accumulation mediates the PDMP-induced alterations. Indeed, an increase in cellular ceramide was measured in PDMP-treated cells. We propose that IBV M is at least in part localized by retrieval mechanisms. Further, ceramide accumulation reveals this cycle by upsetting the balance of anterograde and retrograde traffic and/ or disrupting retention by altering bilayer dynamics.
Abbreviations used in this paper: CGN, cis-Golgi network; GlcCer, glucosylceramide; IBV, infectious bronchitis virus; IC, intermediate compartment; PDMP, d,l-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol; SM, sphingomyelin; VSV, vesicular stomatitis virus.
Address all correspondence to Carolyn E. Machamer, Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, 725 N. Wolfe St., Baltimore, MD 21205. Tel.: (410) 955-1809. Fax: (410) 955-4129. E-mail: carolyn_machamer{at}qmail.bs.jhu.edu

CiteULike
Complore
Connotea
Del.icio.us
Digg
Facebook
Reddit
Technorati
Twitter What's this?
This article has been cited by other articles:
-
Sprocati, T., Ronchi, P., Raimondi, A., Francolini, M., Borgese, N.
(2006). Dynamic and reversible restructuring of the ER induced by PDMP in cultured cells. J. Cell Sci.
119: 3249-3260
[Abstract]
[Full Text]
-
Giussani, P., Maceyka, M., Le Stunff, H., Mikami, A., Lepine, S., Wang, E., Kelly, S., Merrill, A. H. Jr., Milstien, S., Spiegel, S.
(2006). Sphingosine-1-Phosphate Phosphohydrolase Regulates Endoplasmic Reticulum-to-Golgi Trafficking of Ceramide.. Mol. Cell. Biol.
26: 5055-5069
[Abstract]
[Full Text]
-
Nal, B., Chan, C., Kien, F., Siu, L., Tse, J., Chu, K., Kam, J., Staropoli, I., Crescenzo-Chaigne, B., Escriou, N., van der Werf, S., Yuen, K.-Y., Altmeyer, R.
(2005). Differential maturation and subcellular localization of severe acute respiratory syndrome coronavirus surface proteins S, M and E. J. Gen. Virol.
86: 1423-1434
[Abstract]
[Full Text]
-
Schultz, A., Ling, M., Larsson, C.
(2004). Identification of an Amino Acid Residue in the Protein Kinase C C1b Domain Crucial for Its Localization to the Golgi Network. J. Biol. Chem.
279: 31750-31760
[Abstract]
[Full Text]
-
Pettus, B. J., Bielawska, A., Subramanian, P., Wijesinghe, D. S., Maceyka, M., Leslie, C. C., Evans, J. H., Freiberg, J., Roddy, P., Hannun, Y. A., Chalfant, C. E.
(2004). Ceramide 1-Phosphate Is a Direct Activator of Cytosolic Phospholipase A2. J. Biol. Chem.
279: 11320-11326
[Abstract]
[Full Text]
-
Aoki, S., Su, Q., Li, H., Nishikawa, K., Ayukawa, K., Hara, Y., Namikawa, K., Kiryu-Seo, S., Kiyama, H., Wada, K.
(2002). Identification of an Axotomy-Induced Glycosylated Protein, AIGP1, Possibly Involved in Cell Death Triggered by Endoplasmic Reticulum-Golgi Stress. J. Neurosci.
22: 10751-10760
[Abstract]
[Full Text]
-
Holthuis, J. C. M., Pomorski, T., Raggers, R. J., Sprong, H., Van Meer, G.
(2001). The Organizing Potential of Sphingolipids in Intracellular Membrane Transport. Physiol. Rev.
81: 1689-1723
[Abstract]
[Full Text]
-
de Haan, C. A. M., Vennema, H., Rottier, P. J. M.
(2000). Assembly of the Coronavirus Envelope: Homotypic Interactions between the M Proteins. J. Virol.
74: 4967-4978
[Abstract]
[Full Text]
-
Mancini, M., Machamer, C. E., Roy, S., Nicholson, D. W., Thornberry, N. A., Casciola-Rosen, L. A., Rosen, A.
(2000). Caspase-2 Is Localized at the Golgi Complex and Cleaves Golgin-160 during Apoptosis. JCB
149: 603-612
[Abstract]
[Full Text]
-
Fukunaga, T, Nagahama, M, Hatsuzawa, K, Tani, K, Yamamoto, A, Tagaya, M
(2000). Implication of sphingolipid metabolism in the stability of the Golgi apparatus. J. Cell Sci.
113: 3299-3307
[Abstract]
-
Lagace, T. A., Byers, D. M., Cook, H. W., Ridgway, N. D.
(1999). Chinese hamster ovary cells overexpressing the oxysterol binding protein (OSBP) display enhanced synthesis of sphingomyelin in response to 25-hydroxycholesterol. J. Lipid Res.
40: 109-116
[Abstract]
[Full Text]
-
Kok, J. W., Babia, T., Filipeanu, C. M., Nelemans, A., Egea, G., Hoekstra, D.
(1998). PDMP Blocks Brefeldin A-induced Retrograde Membrane Transport from Golgi to ER: Evidence for Involvement of Calcium Homeostasis and Dissociation from Sphingolipid Metabolism. JCB
142: 25-38
[Abstract]
[Full Text]
-
Condrescu, M., Reeves, J. P.
(2001). Inhibition of Sodium-Calcium Exchange by Ceramide and Sphingosine. J. Biol. Chem.
276: 4046-4054
[Abstract]
[Full Text]