title: The basolateral targeting signal of CD147 (EMMPRIN) consists of a single leucine and is not recognized by retinal pigment epithelium.
abstract: 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, because 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, because 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.
authors.affiliation: Margaret M. Dyson Vision Research Institute, Department of Ophthalmology, Weill Medical College of Cornell University, New York, NY 10021, USA.
authors.forename: Ami A
authors.initials: AA
authors.lastname: Deora
authors.forename: Diego
authors.initials: D
authors.lastname: Gravotta
authors.forename: Geri
authors.initials: G
authors.lastname: Kreitzer
authors.forename: Jane
authors.initials: J
authors.lastname: Hu
authors.forename: Dean
authors.initials: D
authors.lastname: Bok
authors.forename: Enrique
authors.initials: E
authors.lastname: Rodriguez-Boulan
mesh_terms: D033961:Adaptor Protein Complex 1 / Q000378:metabolism; D033966:Adaptor Protein Complex beta Subunits / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051926:Basigin; D002460:Cell Line; D016764:Cell Polarity; D017124:Conserved Sequence; D004285:Dogs; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D006207:Half-Life; D006801:Humans; D018374:LLC-PK1 Cells; D007930:Leucine / Q000737:chemistry; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013552:Swine
chemical_list: C439126:AP1B1 protein, human; D033961:Adaptor Protein Complex 1; D033966:Adaptor Protein Complex beta Subunits; D015703:Antigens, CD; C497886:BSG protein, human; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D051926:Basigin; D007930:Leucine