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J. Biol. Chem., Vol. 278, Issue 9, 7325-7334, February 28, 2003
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From the Department of Biochemistry, University of Geneva, Sciences
II, 30 quai E. Ansermet, CH-1211 Geneva 4, Switzerland
Long chain sphingoid bases (LCBs) and their
phosphates (LCBPs) are not only important intermediates in ceramide
biosynthesis but also signaling molecules in the yeast,
Saccharomyces cerevisiae. Their cellular levels, which
control multiple cellular events in response to external and intrinsic
signals, are tightly regulated by coordinated action of metabolic
enzymes such as LCB kinase and LCBP phosphatase. However, little is
known about the mechanisms by which the two enzymes generate
biosynthetic or signaling outputs. It has been shown that the LCBP
phosphatase, Lcb3p, is required for efficient ceramide synthesis from
exogenous LCB. Here we present direct evidence that the major LCB
kinase, Lcb4p, but not the minor kinase, Lcb5p, regulates synthesis of
ceramide from exogenously added LCB. Surprisingly, our biochemical
evidence suggests that the LCBP used for ceramide synthesis must be
generated on the membrane. Our data show that Lcb4p is tightly
associated with membranes and is localized to the endoplasmic reticulum
where it can work in concert with Lcb3p. These results raise the
conceptually attractive possibility that membrane-associated and
cytosolic Lcb4p play distinct roles to differentially generate
biosynthetic and signaling pools of LCBP.
Lcb4p Is a Key Regulator of Ceramide Synthesis from Exogenous
Long Chain Sphingoid Base in Saccharomyces cerevisiae*
*
The work was supported by grants from the Bundesamt
für Bildung und Wissenschaft (EC network Grant HPRN-CT-2000-00077
on Sphingolipids), the Human Frontiers Science Program Organization, and the Swiss National Science Foundation.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.
To whom correspondence should be addressed. Tel.:
41-22-702-6469; Fax: 41-22-702-6465; E-mail:
Howard.Riezman@biochem.unige.ch.
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