Topology of the non-structural rotavirus receptor glycoprotein NS28 in the rough endoplasmic reticulum.
The rotavirus non-structural glycoprotein (NS28), the receptor for the virus core during budding into the lumen of the rough endoplasmic reticulum (RER), is 175 amino acids long and possesses an uncleaved signal sequence and two amino-terminal glycosylation sites. Utilizing one of three potential hydrophobic domains, the protein spans the membrane only once, with the glycosylated amino-terminal region oriented to the luminal side of the ER and the carboxy-terminal region to the cytoplasmic side. To localize sequences involved in translocation of NS28, we constructed a series of mutations in the coding regions for the hydrophobic domains of the protein. Mutant protein products were studied by in vitro translation and by transfection in vivo. In transfected cells, all mutant forms localize to the ER, and none are secreted. In vitro, each of the three hydrophobic domains is able to associate with microsomes. However, glycosylation and proteolysis of wild-type and mutant forms of NS28 indicates that the wild-type protein is anchored in the membrane only by the second hydrophobic domain, leaving approximately 131 residues exposed on the cytoplasmic side for receptor - ligand interaction.
Bergmann
C C
CC
Department of Cellular and Molecular Biology, University of Auckland, New Zealand.
Maass
D
D
Poruchynsky
M S
MS
Atkinson
P H
PH
Bellamy
A R
AR
D000595:Amino Acid Sequence; D001665:Binding Sites; D002872:Chromosome Deletion; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005814:Genes, Viral; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism*; D012401:Rotavirus / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
D008562:Membrane Glycoproteins; D011991:Receptors, Virus; D014764:Viral Proteins