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MBC in Press, published online ahead of print June 23, 2004
Mol. Biol. Cell 10.1091/mbc.E04-01-0058

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Submitted on January 21, 2004
Revised on May 28, 2004
Accepted on June 14, 2004

The Basolateral Targeting Signal of CD147 (EMMPRIN) Consists of a Single Leucine and Is Not Recognized by Retinal Pigment Epithelium

Ami A. Deora*, Diego Gravotta*, Geri Kreitzer{dagger}, Jane Hu{ddagger}, Dean Bok{ddagger}{sect}||, and Enrique Rodriguez-Boulan*¶

*Margaret M. Dyson Vision Research Institute, Department of Ophthalmology, {dagger}Department of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, NY 10021; and {ddagger}Jules Stein Eye Institute, {sect}Brain Research Institute and ||Department of Neurobiology, School of Medicine, University of California, Los Angeles, CA 90095

Monitoring Editor: Juan S. Bonifacino

CD147, a type I integral membrane protein of the immunoglobulin superfamily, exhibits reversed polarity in Retinal Pigment Epithelium (RPE). CD147 is apical in RPE in contrast to its basolateral localization in extraocular epithelia. This elicited our interest in understanding the basolateral sorting signals of CD147 in prototypic Madin-Darby Canine Kidney (MDCK) cells. The cytoplasmic domain of CD147 has basolateral sorting information but is devoid of well-characterized basolateral signals, such as tyrosine and di-leucine motifs. Hence, we carried out systematic site-directed mutagenesis to delineate basolateral targeting information in CD147. Our detailed analysis identified a single leucine (252) as the basolateral targeting motif in the cytoplasmic tail of CD147. Four amino acids (243-246) N-terminal to leucine 252 are also critical basolateral determinants of CD147, since deletion of these amino acids leads to mistargeting of CD147 to the apical membranes. We ruled out the involvement of adaptor complex 1B (AP1B) in the basolateral trafficking of CD147, since LLC-PK1 cells lacking AP1B, target CD147 basolaterally. At variance with MDCK cells, the human RPE cell line ARPE-19 does not distinguish between CD147 (WT) and CD147 with leucine 252 mutated to alanine and targets both proteins apically. Thus, our study identifies an atypical basolateral motif of CD147 which comprises a single leucine and is not recognized by RPE cells. This unusual basolateral sorting signal will be useful in unraveling the specialized sorting machinery of RPE cells.

Corresponding author. E-mail: boulan{at}mail.med.cornell.edu




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