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J. Biol. Chem., Vol. 279, Issue 1, 772-778, January 2, 2004

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FKBP12-Rapamycin-associated Protein or Mammalian Target of Rapamycin (FRAP/mTOR) Localization in the Endoplasmic Reticulum and the Golgi Apparatus^*

Ryan M. Drenan, Xiangyu Liu, Paula G. Bertram, and X. F. Steven Zheng¶

From the Molecular Cell Biology Graduate Program and Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri 63110

FKBP12-rapamycin-associated protein (FRAP) or mammalian target of rapamycin (mTOR) and its effector proteins form a critical signaling pathway that regulates eukaryotic cell growth and proliferation. Although the protein components in this pathway have begun to be identified, little is known about their subcellular localization or the physiological significance of their localization. By immunofluorescence, we find that both endogenous and recombinant FRAP/mTOR proteins show localization predominantly in the endoplasmic reticulum (ER) and the Golgi apparatus. Consistent with this finding, FRAP/mTOR is cofractionated with calnexin, an ER marker protein. Biochemical characterization suggests that FRAP/mTOR is a peripheral ER/Golgi protein with tight membrane association. Finally, we have identified domains of FRAP/mTOR which may mediate its association with the ER and the Golgi apparatus.

Received for publication, June 4, 2003 , and in revised form, October 6, 2003.

^* This work was supported by National Institutes of Health Grants RO1CA099004, RO1CA77668, and RO1GM62817 and by the American Diabetes Association and the Department of Defense (to X. F. S. Z.). 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.

^¶ To whom correspondence should be addressed: Dept. of Pathology and Immunology, Washington University School of Medicine, 660 South Euclid Ave., St. Louis, MO 63110. Tel.: 314-747-1884; Fax: 314-747-1887; E-mail: zheng{at}pathology.wustl.edu.

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