Basolateral sorting signals differ in their ability to redirect apical proteins to the basolateral cell surface.
Polarized sorting of membrane proteins in epithelial cells is mediated by cytoplasmic basolateral signals or by apical signals in the transmembrane or exoplasmic domains. Basolateral signals were generally found to be dominant over apical determinants. We have generated chimeric proteins with the cytoplasmic domain of either the asialoglycoprotein receptor H1 or the transferrin receptor, two basolateral proteins, fused to the transmembrane and exoplasmic segments of aminopeptidase N, an apical protein, and analyzed them in Madin-Darby canine kidney cells. Whereas both cytoplasmic sequences induced endocytosis of the chimeras, only that of the transferrin receptor mediated basolateral expression in steady state. The H1 fusion protein, although still largely sorted to the basolateral side in biosynthetic surface transport, was subsequently resorted to the apical cell surface. We tested whether the difference in sorting between trimeric wild-type H1 and the dimeric aminopeptidase chimera was caused by the number of sorting signals presented in the oligomers. Consistent with this hypothesis, the H1 signal was fully functional in a tetrameric fusion protein with the transmembrane and exoplasmic domains of influenza neuraminidase. The results suggest that basolateral signals per se need not be dominant over apical determinants for steady-state polarity and emphasize an important contribution of the valence of signals in polarized sorting.
Renold
A
A
Biozentrum, University of Basel, CH-4056 Basel, Switzerland.
Cescato
R
R
Beuret
N
N
Vogel
L K
LK
Wahlberg
J M
JM
Brown
J L
JL
Fiedler
K
K
Spiess
M
M
D000595:Amino Acid Sequence; D000818:Animals; D037263:Asialoglycoprotein Receptor; D018826:CD13 Antigens / Q000737:chemistry / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D018076:DNA, Complementary; D004285:Dogs; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000737:chemistry / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism; D011990:Receptors, Transferrin / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction
D037263:Asialoglycoprotein Receptor; D018076:DNA, Complementary; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; D009439:Neuraminidase; D018826:CD13 Antigens