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J. Biol. Chem., Vol. 279, Issue 23, 24561-24568, June 4, 2004

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The T Cell Receptor Chain Alternate Reading Frame Protein (TARP), a Prostate-specific Protein Localized in Mitochondria^*

Hiroshi Maeda, Satoshi Nagata, Curt D. Wolfgang¶, Gary L. Bratthauer||, Tapan K. Bera, and Ira Pastan**

From the Laboratory of Molecular Biology, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20892-4264 and the ^||Department of Gynecologic and Breast Pathology, Armed Forces Institute of Pathology, Washington, D. C. 20306-6000

We previously showed that mRNA encoding TARP (T cell receptor chain alternate reading frame protein) is exclusively expressed in the prostate in males and is up-regulated by androgen in LNCaP cells, an androgen-sensitive prostate cancer cell line. We have now developed an anti-TARP monoclonal antibody named TP1, and show that TARP protein is up-regulated by androgen in both LNCaP and MDA-PCa-2b cells. We used TP1 to determine the subcellular localization of TARP by Western blotting following subcellular fractionation and immunocytochemistry. Both methods showed that TARP is localized in the mitochondria of LNCaP cells, MDA-PCa-2b cells, and PC-3 cells transfected with a TARP-expressing plasmid. We also transfected a plasmid encoding TARP fused to green fluorescent protein into LNCaP, MDA-Pca-2b, and PC-3 cells and confirmed its specific mitochondrial localization in living cells. Fractionation of mitochondria shows that TARP is located in the outer mitochondrial membrane. Immunohistochemistry using a human prostate cancer sample showed that TP1 reacted in a dot-like cytoplasmic pattern consistent with the presence of TARP in mitochondria. These data demonstrate that TARP is the first prostate-specific protein localizing in mitochondria and indicate that TARP, an androgen-regulated protein, may act on mitochondria to carry out its biological functions.

Received for publication, March 4, 2004

^* 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 grants from the Uehara Memorial Foundation and Kanae Foundation for Life & Socio-medical Science.

^¶ Present address: Vanda Pharmaceuticals, 9620 Medical Center Dr., Rockville, MD 20850.

^** To whom correspondence should be addressed: Laboratory of Molecular Biology, NCI, National Institutes of Health, 37 Convent Dr., Room 5106, Bethesda, MD 20892-4264. Tel.: 301-496-4797; Fax: 301-402-1344; E-mail: pastani{at}mail.nih.gov.

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