Original Article

Accumulation of Caveolin in the Endoplasmic Reticulum Redirects the Protein to Lipid Storage Droplets

Correspondence to: Deborah A. Brown, Department of Biochemistry and Cell Biology, SUNY at Stony Brook, Stony Brook, NY 11794-5215. Tel:631-632-8563 Fax:631-632-8575 E-mail:deborah.brown{at}sunysb.edu.

Caveolin-1 is normally localized in plasma membrane caveolae and the Golgi apparatus in mammalian cells. We found three treatments that redirected the protein to lipid storage droplets, identified by staining with the lipophilic dye Nile red and the marker protein ADRP. Caveolin-1 was targeted to the droplets when linked to the ER-retrieval sequence, KKSL, generating Cav–KKSL. Cav–N2, an internal deletion mutant, also accumulated in the droplets, as well as in a Golgi-like structure. Third, incubation of cells with brefeldin A caused caveolin-1 to accumulate in the droplets. This localization persisted after drug washout, showing that caveolin-1 was transported out of the droplets slowly or not at all. Some overexpressed caveolin-2 was also present in lipid droplets. Experimental reduction of cellular cholesteryl ester by 80% did not prevent targeting of Cav–KKSL to the droplets. Cav–KKSL expression did not grossly alter cellular triacylglyceride or cholesteryl levels, although droplet morphology was affected in some cells. These data suggest that accumulation of caveolin-1 to unusually high levels in the ER causes targeting to lipid droplets, and that mechanisms must exist to ensure the rapid exit of newly synthesized caveolin-1 from the ER to avoid this fate.

Key Words: caveolae, brefeldin A, retrograde transport, triacylglycerol, cholesteryl ester

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Barr, D. J., Ostermeyer-Fay, A. G., Matundan, R. A., Brown, D. A. (2008). Clathrin-independent endocytosis of ErbB2 in geldanamycin-treated human breast cancer cells. J. Cell Sci. 121: 3155-3166 [Abstract] [Full Text]  
• Anderson, N., Borlak, J. (2008). Molecular Mechanisms and Therapeutic Targets in Steatosis and Steatohepatitis. Pharmacol. Rev. 60: 311-357 [Abstract] [Full Text]  
• Lee, C.-J., Lin, H.-R., Liao, C.-L., Lin, Y.-L. (2008). Cholesterol Effectively Blocks Entry of Flavivirus. J. Virol. 82: 6470-6480 [Abstract] [Full Text]  
• Zehmer, J. K., Bartz, R., Liu, P., Anderson, R. G. W. (2008). Identification of a novel N-terminal hydrophobic sequence that targets proteins to lipid droplets. J. Cell Sci. 121: 1852-1860 [Abstract] [Full Text]  
• Garg, A., Agarwal, A. K. (2008). Caveolin-1: A New Locus for Human Lipodystrophy. J. Clin. Endocrinol. Metab. 93: 1183-1185 [Full Text]  
• Pilch, P. F., Souto, R. P., Liu, L., Jedrychowski, M. P., Berg, E. A., Costello, C. E., Gygi, S. P. (2007). Cellular spelunking: exploring adipocyte caveolae. J. Lipid Res. 48: 2103-2111 [Abstract] [Full Text]  
• Predescu, S. A., Predescu, D. N., Malik, A. B. (2007). Molecular determinants of endothelial transcytosis and their role in endothelial permeability. Am. J. Physiol. Lung Cell. Mol. Physiol. 293: L823-L842 [Abstract] [Full Text]  
• Tsuiki, E., Fujita, A., Ohsaki, Y., Cheng, J., Irie, T., Yoshikawa, K., Senoo, H., Mishima, K., Kitaoka, T., Fujimoto, T. (2007). All-trans-Retinol Generated by Rhodopsin Photobleaching Induces Rapid Recruitment of TIP47 to Lipid Droplets in the Retinal Pigment Epithelium. IOVS 48: 2858-2867 [Abstract] [Full Text]  
• Fujimoto, Y., Itabe, H., Kinoshita, T., Homma, K. J., Onoduka, J., Mori, M., Yamaguchi, S., Makita, M., Higashi, Y., Yamashita, A., Takano, T. (2007). Involvement of ACSL in local synthesis of neutral lipids in cytoplasmic lipid droplets in human hepatocyte HuH7. J. Lipid Res. 48: 1280-1292 [Abstract] [Full Text]  
• Sun, X.-H., Flynn, D. C., Castranova, V., Millecchia, L. L., Beardsley, A. R., Liu, J. (2007). Identification of a Novel Domain at the N Terminus of Caveolin-1 That Controls Rear Polarization of the Protein and Caveolae Formation. J. Biol. Chem. 282: 7232-7241 [Abstract] [Full Text]  
• Sturmey, R G, O'Toole, P J, Leese, H J (2006). Fluorescence resonance energy transfer analysis of mitochondrial:lipid association in the porcine oocyte.. Reproduction 132: 829-837 [Abstract] [Full Text]  
• Frank, P. G., Cheung, M. W.-C., Pavlides, S., Llaverias, G., Park, D. S., Lisanti, M. P. (2006). Caveolin-1 and regulation of cellular cholesterol homeostasis. Am. J. Physiol. Heart Circ. Physiol. 291: H677-H686 [Abstract] [Full Text]  
• Andersson, L., Bostrom, P., Ericson, J., Rutberg, M., Magnusson, B., Marchesan, D., Ruiz, M., Asp, L., Huang, P., Frohman, M. A., Boren, J., Olofsson, S.-O. (2006). PLD1 and ERK2 regulate cytosolic lipid droplet formation. J. Cell Sci. 119: 2246-2257 [Abstract] [Full Text]  
• Manninen, A., Verkade, P., Le Lay, S., Torkko, J., Kasper, M., Fullekrug, J., Simons, K. (2005). Caveolin-1 Is Not Essential for Biosynthetic Apical Membrane Transport. Mol. Cell. Biol. 25: 10087-10096 [Abstract] [Full Text]  
• Robenek, H., Robenek, M. J., Buers, I., Lorkowski, S., Hofnagel, O., Troyer, D., Severs, N. J. (2005). Lipid Droplets Gain PAT Family Proteins by Interaction with Specialized Plasma Membrane Domains. J. Biol. Chem. 280: 26330-26338 [Abstract] [Full Text]  
• Ozeki, S., Cheng, J., Tauchi-Sato, K., Hatano, N., Taniguchi, H., Fujimoto, T. (2005). Rab18 localizes to lipid droplets and induces their close apposition to the endoplasmic reticulum-derived membrane. J. Cell Sci. 118: 2601-2611 [Abstract] [Full Text]  
• Robenek, H., Robenek, M. J., Troyer, D. (2005). PAT family proteins pervade lipid droplet cores. J. Lipid Res. 46: 1331-1338 [Abstract] [Full Text]  
• Wolins, N. E., Quaynor, B. K., Skinner, J. R., Schoenfish, M. J., Tzekov, A., Bickel, P. E. (2005). S3-12, Adipophilin, and TIP47 Package Lipid in Adipocytes. J. Biol. Chem. 280: 19146-19155 [Abstract] [Full Text]  
• Heuser, J. (2005). Deep-etch EM reveals that the early poxvirus envelope is a single membrane bilayer stabilized by a geodetic "honeycomb" surface coat. JCB 169: 269-283 [Abstract] [Full Text]  
• Meadows, J. W., Pitzer, B., Brockman, D. E., Myatt, L. (2005). Expression and Localization of Adipophilin and Perilipin in Human Fetal Membranes: Association with Lipid Bodies and Enzymes Involved in Prostaglandin Synthesis. J. Clin. Endocrinol. Metab. 90: 2344-2350 [Abstract] [Full Text]  
• Pol, A., Martin, S., Fernandez, M. A., Ingelmo-Torres, M., Ferguson, C., Enrich, C., Parton, R. G. (2005). Cholesterol and Fatty Acids Regulate Dynamic Caveolin Trafficking through the Golgi Complex and between the Cell Surface and Lipid Bodies. Mol. Biol. Cell 16: 2091-2105 [Abstract] [Full Text]  
• Puljak, L., Pagliassotti, M. J, Wei, Y., Qadri, I., Parameswara, V., Esser, V., Fitz, J. G., Kilic, G. (2005). Inhibition of cellular responses to insulin in a rat liver cell line. A role for PKC in insulin resistance. J. Physiol. 563: 471-482 [Abstract] [Full Text]  
• Brasaemle, D. L., Dolios, G., Shapiro, L., Wang, R. (2004). Proteomic Analysis of Proteins Associated with Lipid Droplets of Basal and Lipolytically Stimulated 3T3-L1 Adipocytes. J. Biol. Chem. 279: 46835-46842 [Abstract] [Full Text]  
• Subramanian, V., Garcia, A., Sekowski, A., Brasaemle, D. L. (2004). Hydrophobic sequences target and anchor perilipin A to lipid droplets. J. Lipid Res. 45: 1983-1991 [Abstract] [Full Text]  
• Llorente, A., de Marco, M. C., Alonso, M. A. (2004). Caveolin-1 and MAL are located on prostasomes secreted by the prostate cancer PC-3 cell line. J. Cell Sci. 117: 5343-5351 [Abstract] [Full Text]  
• Cohen, A. W., Hnasko, R., Schubert, W., Lisanti, M. P. (2004). Role of Caveolae and Caveolins in Health and Disease. Physiol. Rev. 84: 1341-1379 [Abstract] [Full Text]  
• Ren, X., Ostermeyer, A. G., Ramcharan, L. T., Zeng, Y., Lublin, D. M., Brown, D. A. (2004). Conformational Defects Slow Golgi Exit, Block Oligomerization, and Reduce Raft Affinity of Caveolin-1 Mutant Proteins. Mol. Biol. Cell 15: 4556-4567 [Abstract] [Full Text]  
• Imanishi, Y., Gerke, V., Palczewski, K. (2004). Retinosomes: new insights into intracellular managing of hydrophobic substances in lipid bodies. JCB 166: 447-453 [Abstract] [Full Text]  
• Umlauf, E., Csaszar, E., Moertelmaier, M., Schuetz, G. J., Parton, R. G., Prohaska, R. (2004). Association of Stomatin with Lipid Bodies. J. Biol. Chem. 279: 23699-23709 [Abstract] [Full Text]  
• Cohen, A. W., Razani, B., Schubert, W., Williams, T. M., Wang, X. B., Iyengar, P., Brasaemle, D. L., Scherer, P. E., Lisanti, M. P. (2004). Role of Caveolin-1 in the Modulation of Lipolysis and Lipid Droplet Formation. Diabetes 53: 1261-1270 [Abstract] [Full Text]  
• Wadehra, M., Goodglick, L., Braun, J. (2004). The Tetraspan Protein EMP2 Modulates the Surface Expression of Caveolins and Glycosylphosphatidyl Inositol-linked Proteins. Mol. Biol. Cell 15: 2073-2083 [Abstract] [Full Text]  
• Larigauderie, G., Furman, C., Jaye, M., Lasselin, C., Copin, C., Fruchart, J.-C., Castro, G., Rouis, M. (2004). Adipophilin Enhances Lipid Accumulation and Prevents Lipid Efflux From THP-1 Macrophages: Potential Role in Atherogenesis. Arterioscler. Thromb. Vasc. Bio. 24: 504-510 [Abstract] [Full Text]  
• Kamisaka, Y., Noda, N., Yamaoka, M. (2004). Appearance of Smaller Lipid Bodies and Protein Kinase Activation in the Lipid Body Fraction Are Induced by an Increase in the Nitrogen Source in the Mortierella Fungus. J Biochem 135: 269-276 [Abstract] [Full Text]  
• Ostermeyer, A. G., Ramcharan, L. T., Zeng, Y., Lublin, D. M., Brown, D. A. (2004). Role of the hydrophobic domain in targeting caveolin-1 to lipid droplets. JCB 164: 69-78 [Abstract] [Full Text]  
• Pol, A., Martin, S., Fernandez, M. A., Ferguson, C., Carozzi, A., Luetterforst, R., Enrich, C., Parton, R. G. (2004). Dynamic and Regulated Association of Caveolin with Lipid Bodies: Modulation of Lipid Body Motility and Function by a Dominant Negative Mutant. Mol. Biol. Cell 15: 99-110 [Abstract] [Full Text]  
• Hnasko, R., Lisanti, M. P. (2003). The Biology of Caveolae: Lessons from Caveolin Knockout Mice and Implications for Human Disease. Mol. Interv. 3: 445-464 [Abstract] [Full Text]  
• Caldas, H., Herman, G. E. (2003). NSDHL, an enzyme involved in cholesterol biosynthesis, traffics through the Golgi and accumulates on ER membranes and on the surface of lipid droplets. Hum Mol Genet 12: 2981-2991 [Abstract] [Full Text]  
• Dagher, G., Donne, N., Klein, C., Ferre, P., Dugail, I. (2003). HDL-mediated cholesterol uptake and targeting to lipid droplets in adipocytes. J. Lipid Res. 44: 1811-1820 [Abstract] [Full Text]  
• Ohashi, M., Mizushima, N., Kabeya, Y., Yoshimori, T. (2003). Localization of Mammalian NAD(P)H Steroid Dehydrogenase-like Protein on Lipid Droplets. J. Biol. Chem. 278: 36819-36829 [Abstract] [Full Text]  
• Hayashi, T., Su, T.-P. (2003). {sigma}-1 Receptors ({sigma}1 Binding Sites) Form Raft-Like Microdomains and Target Lipid Droplets on the Endoplasmic Reticulum: Roles in Endoplasmic Reticulum Lipid Compartmentalization and Export. J. Pharmacol. Exp. Ther. 306: 718-725 [Abstract] [Full Text]  
• Marchesan, D., Rutberg, M., Andersson, L., Asp, L., Larsson, T., Boren, J., Johansson, B. R., Olofsson, S.-O. (2003). A Phospholipase D-dependent Process Forms Lipid Droplets Containing Caveolin, Adipocyte Differentiation-related Protein, and Vimentin in a Cell-free System. J. Biol. Chem. 278: 27293-27300 [Abstract] [Full Text]  
• DiDonato, D., Brasaemle, D. L. (2003). Fixation Methods for the Study of Lipid Droplets by Immunofluorescence Microscopy. J. Histochem. Cytochem. 51: 773-780 [Abstract] [Full Text]  
• Souto, R. P., Vallega, G., Wharton, J., Vinten, J., Tranum-Jensen, J., Pilch, P. F. (2003). Immunopurification and Characterization of Rat Adipocyte Caveolae Suggest Their Dissociation from Insulin Signaling. J. Biol. Chem. 278: 18321-18329 [Abstract] [Full Text]  
• Shi, S. T., Lee, K.-J., Aizaki, H., Hwang, S. B., Lai, M. M. C. (2003). Hepatitis C Virus RNA Replication Occurs on a Detergent-Resistant Membrane That Cofractionates with Caveolin-2. J. Virol. 77: 4160-4168 [Abstract] [Full Text]  
• Garcia, A., Sekowski, A., Subramanian, V., Brasaemle, D. L. (2003). The Central Domain Is Required to Target and Anchor Perilipin A to Lipid Droplets. J. Biol. Chem. 278: 625-635 [Abstract] [Full Text]  
• Tauchi-Sato, K., Ozeki, S., Houjou, T., Taguchi, R., Fujimoto, T. (2002). The Surface of Lipid Droplets Is a Phospholipid Monolayer with a Unique Fatty Acid Composition. J. Biol. Chem. 277: 44507-44512 [Abstract] [Full Text]  
• Wu, D., Terrian, D. M. (2002). Regulation of Caveolin-1 Expression and Secretion by a Protein Kinase Cepsilon Signaling Pathway in Human Prostate Cancer Cells. J. Biol. Chem. 277: 40449-40455 [Abstract] [Full Text]  
• Razani, B., Woodman, S. E., Lisanti, M. P. (2002). Caveolae: From Cell Biology to Animal Physiology. Pharmacol. Rev. 54: 431-467 [Abstract] [Full Text]  
• Miura, S., Gan, J.-W., Brzostowski, J., Parisi, M. J., Schultz, C. J., Londos, C., Oliver, B., Kimmel, A. R. (2002). Functional Conservation for Lipid Storage Droplet Association among Perilipin, ADRP, and TIP47 (PAT)-related Proteins in Mammals, Drosophila, and Dictyostelium. J. Biol. Chem. 277: 32253-32257 [Abstract] [Full Text]  
• Garver, W. S., Krishnan, K., Gallagos, J. R., Michikawa, M., Francis, G. A., Heidenreich, R. A. (2002). Niemann-Pick C1 protein regulates cholesterol transport to the trans-Golgi network and plasma membrane caveolae. J. Lipid Res. 43: 579-589 [Abstract] [Full Text]  
• Razani, B., Wang, X. B., Engelman, J. A., Battista, M., Lagaud, G., Zhang, X. L., Kneitz, B., Hou, H. Jr., Christ, G. J., Edelmann, W., Lisanti, M. P. (2002). Caveolin-2-Deficient Mice Show Evidence of Severe Pulmonary Dysfunction without Disruption of Caveolae. Mol. Cell. Biol. 22: 2329-2344 [Abstract] [Full Text]  
• Wu, D., Foreman, T. L., Gregory, C. W., McJilton, M. A., Wescott, G. G., Ford, O. H., Alvey, R. F., Mohler, J. L., Terrian, D. M. (2002). Protein Kinase C{epsilon} Has the Potential to Advance the Recurrence of Human Prostate Cancer. Cancer Res. 62: 2423-2429 [Abstract] [Full Text]  
• Hope, R. G., Murphy, D. J., McLauchlan, J. (2002). The Domains Required to Direct Core Proteins of Hepatitis C Virus and GB Virus-B to Lipid Droplets Share Common Features with Plant Oleosin Proteins. J. Biol. Chem. 277: 4261-4270 [Abstract] [Full Text]  
• Thomsen, P., Roepstorff, K., Stahlhut, M., van Deurs, B. (2002). Caveolae Are Highly Immobile Plasma Membrane Microdomains, Which Are not Involved in Constitutive Endocytic Trafficking. Mol. Biol. Cell 13: 238-250 [Abstract] [Full Text]  
• van Meer, G. (2001). Caveolin, Cholesterol, and Lipid Droplets?. JCB 152: F29-F34 [Abstract] [Full Text]  
• Razani, B., Combs, T. P., Wang, X. B., Frank, P. G., Park, D. S., Russell, R. G., Li, M., Tang, B., Jelicks, L. A., Scherer, P. E., Lisanti, M. P. (2002). Caveolin-1-deficient Mice Are Lean, Resistant to Diet-induced Obesity, and Show Hypertriglyceridemia with Adipocyte Abnormalities. J. Biol. Chem. 277: 8635-8647 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents