title: Polarized apical targeting directed by the signal/anchor region of simian virus 5 hemagglutinin-neuraminidase.
abstract: To examine the possibility of independent cytoplasmic/transmembrane domain-based apical sorting, we have investigated paramyxovirus SV5 hemagglutinin-neuraminidase (HN), a type II membrane protein with a small N-terminal signal/anchor region. In SV5-infected Madin-Darby canine kidney (MDCK) cells, >90% of HN is found on the apical surface. We have expressed chimeric proteins in which the N terminus of HN, including its signal/anchor region, is attached to a (normally cytosolic) reporter pyruvate kinase (PK). PK itself expressed immediately downstream from a cleavable signal peptide was converted to a 58-kDa N-linked glycosylated form, which was secreted predominantly (80%) to the basolateral surface of MDCK cells. By contrast, stably expressed PK chimeras, now anchored as type II membrane proteins with either the first 48 or 72 amino acids of HN, received similar N-linked glycosylation, yet exhibited polarized transport with a preferentially (75%) apical distribution. These results suggest that the N-terminal signal/anchor region of HN contains independent sorting information for apical specific targeting in MDCK cells.
authors.affiliation: Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35209, USA.
authors.forename: X F
authors.initials: XF
authors.lastname: Huang
authors.forename: R W
authors.initials: RW
authors.lastname: Compans
authors.forename: S
authors.initials: S
authors.lastname: Chen
authors.forename: R A
authors.initials: RA
authors.lastname: Lamb
authors.forename: P
authors.initials: P
authors.lastname: Arvan
mesh_terms: D000818:Animals; D002460:Cell Line; D004285:Dogs; D005089:Exocytosis; D015707:HN Protein / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D021382:Protein Sorting Signals / Q000378:metabolism*; D011770:Pyruvate Kinase / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D018108:Respirovirus / Q000201:enzymology / Q000378:metabolism*
chemical_list: D015707:HN Protein; D008565:Membrane Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D011770:Pyruvate Kinase