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J. Biol. Chem., Vol. 279, Issue 36, 37798-37807, September 3, 2004

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Mammalian Wax Biosynthesis

II. EXPRESSION CLONING OF WAX SYNTHASE cDNAs ENCODING A MEMBER OF THE ACYLTRANSFERASE ENZYME FAMILY^*

Jeffrey B. Cheng and David W. Russell

From the Department of Molecular Genetics, The University of Texas Southwestern Medical Center, Dallas, Texas 75390

Wax monoesters are synthesized by the esterification of fatty alcohols and fatty acids. A mammalian enzyme that catalyzes this reaction has not been isolated. We used expression cloning to identify cDNAs encoding a wax synthase in the mouse preputial gland. The wax synthase gene is located on the X chromosome and encodes a member of the acyltransferase family of enzymes that synthesize neutral lipids. Expression of wax synthase in cultured cells led to the formation of wax monoesters from straight chain saturated, unsaturated, and polyunsaturated fatty alcohols and acids. Polyisoprenols also were incorporated into wax monoesters by the enzyme. The wax synthase had little or no ability to synthesize cholesteryl esters, diacylglycerols, or triacylglycerols, whereas other acyltransferases, including the acyl-CoA:monoacylglycerol acyltransferase 1 and 2 enzymes and the acyl-CoA:diacylglycerol acyltransferase 1 and 2 enzymes, exhibited modest wax monoester synthesis activities. Confocal light microscopy indicated that the wax synthase was localized in membranes of the endoplasmic reticulum. Wax synthase mRNA was abundant in tissues rich in sebaceous glands such as the preputial gland and eyelid and was present at lower levels in other tissues. Coexpression of cDNAs specifying fatty acyl-CoA reductase 1 and wax synthase led to the synthesis of wax monoesters. The data suggest that wax monoester synthesis in mammals involves a two step biosynthetic pathway catalyzed by fatty acyl-CoA reductase and wax synthase enzymes.

Received for publication, June 4, 2004 , and in revised form, June 24, 2004.

^* This work was supported in part by National Institutes of Health Grant HL20948 (to D. W. R.) and Robert A. Welch Foundation Grant I-0971 (to D. W. R.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Supported by National Institutes of Health Medical Scientist Training Grant GM08014.

To whom correspondence should be addressed: Dept. of Molecular Genetics, UT Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9046. Tel.: 214-648-2007; Fax: 214-648-6899; E-mail: david.russell{at}utsouthwestern.edu.

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