Transient translocation of the cytoplasmic (endo) domain of a type I membrane glycoprotein into cellular membranes.
The E2 glycoprotein of the alphavirus Sindbis is a typical type I membrane protein with a single membrane spanning domain and a cytoplasmic tail (endo domain) containing 33 amino acids. The carboxyl terminal domain of the tail has been implicated as (a) attachment site for nucleocapsid protein, and (b) signal sequence for integration of the other alpha-virus membrane proteins 6K and E1. These two functions require that the carboxyl terminus be exposed in the cell cytoplasm (a) and exposed in the lumen of the endoplasmic reticulum (b). We have investigated the orientation of this glycoprotein domain with respect to cell membranes by substituting a tyrosine for the normally occurring serine, four amino acids upstream of the carboxyl terminus. Using radioiodination of this tyrosine as an indication of the exposure of the glycoprotein tail, we have provided evidence that this domain is initially translocated into a membrane and is returned to the cytoplasm after export from the ER. This is the first demonstration of such a transient translocation of a single domain of an integral membrane protein and this rearrangement explains some important aspects of alphavirus assembly.
Liu
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Cell Research Institute, University of Texas, Austin 78713-7640.
Brown
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D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D066298:In Vitro Techniques; D008562:Membrane Glycoproteins / Q000378:metabolism* / Q000648:ultrastructure; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D012845:Sindbis Virus; D014759:Viral Envelope Proteins / Q000378:metabolism* / Q000648:ultrastructure
D008562:Membrane Glycoproteins; D009838:Oligodeoxyribonucleotides; D014759:Viral Envelope Proteins