JBC Ideal method for primary cell transfection

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M007721200 on September 26, 2000

J. Biol. Chem., Vol. 275, Issue 50, 39793-39798, December 15, 2000
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
275/50/39793    most recent
M007721200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sawai, H.
Right arrow Articles by Hannun, Y. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sawai, H.
Right arrow Articles by Hannun, Y. A.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Identification of ISC1 (YER019w) as Inositol Phosphosphingolipid Phospholipase C in Saccharomyces cerevisiae*

Hirofumi SawaiDagger §, Yasuo OkamotoDagger , Chiara LubertoDagger , Cungui Mao||, Alicja BielawskaDagger , Naochika Domae§, and Yusuf A. HannunDagger **

From the Dagger  Department of Biochemistry and Molecular Biology and the || Ralph H. Johnson Veterans Administration Hospital and the Department of Medicine, Medical University of South Carolina, Charleston, South Carolina 29425 and the § Department of Medicine, Osaka Dental University, 8-1 Kuzuhahanazonocho, Hirakata, Osaka 573, Japan

Sphingolipids have emerged as novel bioactive mediators in eukaryotic cells including yeast. It has been proposed that sphingomyelin (SM) hydrolysis and the concomitant generation of ceramide are involved in various stress responses in mammalian cells. The yeast Saccharomyces cerevisiae has inositol phosphosphingolipids (IPS) instead of SM and glycolipids, and synthesis of IPS is indispensable to its growth. Although the genes responsible for the synthesis of IPS have been identified, the gene(s) for the degradation of IPS has not been reported. Here we show that ISC1 (YER019w), which has homology to bacterial neutral sphingomyelinase (SMase), encodes IPS phospholipase C (IPS-PLC). First, we observed that overexpression of ISC1 greatly increased neutral SMase activity, and this activity was dependent on the presence of phosphatidylserine. Cells deleted in ISC1 demonstrated negligible neutral SMase activity. Because yeast cells have IPS instead of SM, we investigated whether IPS are the physiologic substrates of this enzyme. Lysates of ISC1-overexpressing cells demonstrated very high PLC activities on IPS. Deletion of ISC1 eliminated endogenous IPS-PLC activities. Labeling yeast cells with [3H]dihydrosphingosine showed that IPS were increased in the deletion mutant cells. This study identifies the first enzyme involved in catabolism of complex sphingolipids in S. cerevisiae.

* This work was supported in part by National Institutes of Health Grant GM43825 (to Y. A. H.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of the Merck Company Foundation and Banyu Fellowship Awards in Lipid Metabolism and Atherosclerosis.

** To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, Medical University of South Carolina, 173 Ashley Ave., Charleston, SC 29425. Tel.: 843-792-4321; Fax: 843-792-4322, E-mail: hannun@musc.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 J. Lipid Res.Home page
R. C. Dickson
Thematic Review Series: Sphingolipids. New insights into sphingolipid metabolism and function in budding yeast
J. Lipid Res., May 1, 2008; 49(5): 909 - 921.
[Abstract] [Full Text] [PDF]

Home page
 J. Lipid Res.Home page
N. Matmati and Y. A. Hannun
Thematic Review Series: Sphingolipids. ISC1 (inositol phosphosphingolipid-phospholipase C), the yeast homologue of neutral sphingomyelinases
J. Lipid Res., May 1, 2008; 49(5): 922 - 928.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
T. Almeida, M. Marques, D. Mojzita, M. A. Amorim, R. D. Silva, B. Almeida, P. Rodrigues, P. Ludovico, S. Hohmann, P. Moradas-Ferreira, et al.
Isc1p Plays a Key Role in Hydrogen Peroxide Resistance and Chronological Lifespan through Modulation of Iron Levels and Apoptosis
Mol. Biol. Cell, March 1, 2008; 19(3): 865 - 876.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
M. Umemura, M. Fujita, T. Yoko-o, A. Fukamizu, and Y. Jigami
Saccharomyces cerevisiae CWH43 Is Involved in the Remodeling of the Lipid Moiety of GPI Anchors to Ceramides
Mol. Biol. Cell, November 1, 2007; 18(11): 4304 - 4316.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
R. Rhome, T. McQuiston, T. Kechichian, A. Bielawska, M. Hennig, M. Drago, G. Morace, C. Luberto, and M. Del Poeta
Biosynthesis and Immunogenicity of Glucosylceramide in Cryptococcus neoformans and Other Human Pathogens
Eukaryot. Cell, October 1, 2007; 6(10): 1715 - 1726.
[Full Text] [PDF]

Home page
 Infect. Immun.Home page
J. M. Shea, T. B. Kechichian, C. Luberto, and M. Del Poeta
The cryptococcal enzyme inositol phosphosphingolipid-phospholipase C confers resistance to the antifungal effects of macrophages and promotes fungal dissemination to the central nervous system.
Infect. Immun., October 1, 2006; 74(10): 5977 - 5988.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
M. Tabuchi, A. Audhya, A. B. Parsons, C. Boone, and S. D. Emr
The Phosphatidylinositol 4,5-Biphosphate and TORC2 Binding Proteins Slm1 and Slm2 Function in Sphingolipid Regulation
Mol. Cell. Biol., August 1, 2006; 26(15): 5861 - 5875.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
M. Osterberg, H. Kim, J. Warringer, K. Melen, A. Blomberg, and G. von Heijne
Phenotypic effects of membrane protein overexpression in Saccharomyces cerevisiae
PNAS, July 25, 2006; 103(30): 11148 - 11153.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
O. Krut, K. Wiegmann, H. Kashkar, B. Yazdanpanah, and M. Kronke
Novel Tumor Necrosis Factor-responsive Mammalian Neutral Sphingomyelinase-3 Is a C-tail-anchored Protein
J. Biol. Chem., May 12, 2006; 281(19): 13784 - 13793.
[Abstract] [Full Text] [PDF]

Home page
 GlycobiologyHome page
M. Pittet, D. Uldry, M. Aebi, and A. Conzelmann
The N-glycosylation defect of cwh8{Delta} yeast cells causes a distinct defect in sphingolipid biosynthesis
Glycobiology, February 1, 2006; 16(2): 155 - 164.
[Abstract] [Full Text] [PDF]

Home page
 Eukaryot CellHome page
S. Zink, C. Mehlgarten, H. K. Kitamoto, J. Nagase, D. Jablonowski, R. C. Dickson, M. J. R. Stark, and R. Schaffrath
Mannosyl-Diinositolphospho-Ceramide, the Major Yeast Plasma Membrane Sphingolipid, Governs Toxicity of Kluyveromyces lactis Zymocin
Eukaryot. Cell, May 1, 2005; 4(5): 879 - 889.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. Vaena de Avalos, X. Su, M. Zhang, Y. Okamoto, W. Dowhan, and Y. A. Hannun
The Phosphatidylglycerol/Cardiolipin Biosynthetic Pathway Is Required for the Activation of Inositol Phosphosphingolipid Phospholipase C, Isc1p, during Growth of Saccharomyces cerevisiae
J. Biol. Chem., February 25, 2005; 280(8): 7170 - 7177.
[Abstract] [Full Text] [PDF]

Home page
 ANN BOT (LOND)Home page
T. M. DUNN, D. V. LYNCH, L. V. MICHAELSON, and J. A. NAPIER
A Post-genomic Approach to Understanding Sphingolipid Metabolism in Arabidopsis thaliana
Ann. Bot., May 1, 2004; 93(5): 483 - 497.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
S. V. de Avalos, Y. Okamoto, and Y. A. Hannun
Activation and Localization of Inositol Phosphosphingolipid Phospholipase C, Isc1p, to the Mitochondria during Growth of Saccharomyces cerevisiae
J. Biol. Chem., March 19, 2004; 279(12): 11537 - 11545.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Luberto, M. J. Stonehouse, E. A. Collins, N. Marchesini, S. El-Bawab, A. I. Vasil, M. L. Vasil, and Y. A. Hannun
Purification, Characterization, and Identification of a Sphingomyelin Synthase from Pseudomonas aeruginosa: PlcH IS A MULTIFUNCTIONAL ENZYME
J. Biol. Chem., August 29, 2003; 278(35): 32733 - 32743.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. A. Cowart, Y. Okamoto, F. R. Pinto, J. L. Gandy, J. S. Almeida, and Y. A. Hannun
Roles for Sphingolipid Biosynthesis in Mediation of Specific Programs of the Heat Stress Response Determined through Gene Expression Profiling
J. Biol. Chem., August 8, 2003; 278(32): 30328 - 30338.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
N. Marchesini, C. Luberto, and Y. A. Hannun
Biochemical Properties of Mammalian Neutral Sphingomyelinase2 and Its Role in Sphingolipid Metabolism
J. Biol. Chem., April 11, 2003; 278(16): 13775 - 13783.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
U. D. Epple, E.-L. Eskelinen, and M. Thumm
Intravacuolar Membrane Lysis in Saccharomyces cerevisiae. DOES VACUOLAR TARGETING OF Cvt17/Aut5p AFFECT ITS FUNCTION?
J. Biol. Chem., February 28, 2003; 278(10): 7810 - 7821.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
Y. Okamoto, S. Vaena de Avalos, and Y. A. Hannun
Structural Requirements for Selective Binding of ISC1 to Anionic Phospholipids
J. Biol. Chem., November 22, 2002; 277(48): 46470 - 46477.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Cell. Biol.Home page
M. Zumbansen and W. Stoffel
Neutral Sphingomyelinase 1 Deficiency in the Mouse Causes No Lipid Storage Disease
Mol. Cell. Biol., June 1, 2002; 22(11): 3633 - 3638.
[Abstract] [Full Text] [PDF]

Home page
 Physiol. Rev.Home page
J. C. M. Holthuis, T. Pomorski, R. J. Raggers, H. Sprong, and G. Van Meer
The Organizing Potential of Sphingolipids in Intracellular Membrane Transport
Physiol Rev, October 1, 2001; 81(4): 1689 - 1723.
[Abstract] [Full Text] [PDF]

Home page
 GeneticsHome page
A. Feoktistova, P. Magnelli, C. Abeijon, P. Perez, R. L. Lester, R. C. Dickson, and K. L. Gould
Coordination Between Fission Yeast Glucan Formation and Growth Requires a Sphingolipase Activity
Genetics, August 1, 2001; 158(4): 1397 - 1411.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2000 by the American Society for Biochemistry and Molecular Biology.