Transmembrane domain of influenza virus neuraminidase, a type II protein, possesses an apical sorting signal in polarized MDCK cells.
The influenza virus neuraminidase (NA), a type II transmembrane protein, is directly transported to the apical plasma membrane in polarized MDCK cells. By using deletion mutants and chimeric constructs of influenza virus NA with the human transferrin receptor, a type II basolateral transmembrane protein, we investigated the location of the apical sorting signal of influenza virus NA. When these mutant and chimeric proteins were expressed in stably transfected polarized MDCK cells, the transmembrane domain of NA, and not the cytoplasmic tail, provided a determinant for apical targeting in polarized MDCK cells and this transmembrane signal was sufficient for sorting and transport of the ectodomain of a reporter protein (transferrin receptor) directly to the apical plasma membrane of polarized MDCK cells. In addition, by using differential detergent extraction, we demonstrated that influenza virus NA and the chimeras which were transported to the apical plasma membrane also became insoluble in Triton X-100 but soluble in octylglucoside after extraction from MDCK cells during exocytic transport. These data indicate that the transmembrane domain of NA provides the determinant(s) both for apical transport and for association with Triton X-100-insoluble lipids.
Kundu
A
A
Jonsson Comprehensive Cancer Center, Department of Microbiology and Immunology, University of California at Los Angeles 90095-1747, USA.
Avalos
R T
RT
Sanderson
C M
CM
Nayak
D P
DP
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000502:physiology; D004279:DNA, Viral / Q000737:chemistry; D003902:Detergents; D004285:Dogs; D005089:Exocytosis; D005960:Glucosides; D006801:Humans; D007668:Kidney; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009439:Neuraminidase / Q000096:biosynthesis / Q000737:chemistry / Q000502:physiology*; D017830:Octoxynol; D009975:Orthomyxoviridae / Q000201:enzymology*; D011990:Receptors, Transferrin / Q000096:biosynthesis; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017384:Sequence Deletion; D015398:Signal Transduction
D004279:DNA, Viral; D003902:Detergents; D005960:Glucosides; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; C018619:octyl-beta-D-glucoside; D017830:Octoxynol; D009439:Neuraminidase