The E1 glycoprotein of an avian coronavirus is targeted to the cis Golgi complex.
It was previously reported that the E1 protein of an avian coronavirus was targeted to the juxtanuclear region in COS cells expressing the protein from cloned cDNA, suggesting that the protein contains information for targeting to the Golgi complex. The first of three membrane-spanning domains was required for intracellular targeting, because a mutant E1 (delta m1,2) lacking this domain was delivered to the plasma membrane. We have used immunoelectron microscopy to localize the wild-type E1 protein within Golgi elements of COS cells and AtT-20 cells expressing these proteins from recombinant vaccinia vectors. By immunoperoxidase and immunogold labeling, the wild-type E1 protein was localized to one or two cisternae located on one side of the Golgi stack that could be identified as the cis side in AtT-20 cells. In contrast, the mutant E1 protein was detected in all cisternae across the stack as well as at the plasma membrane. When the E1 proteins were immunoprecipitated and subjected to digestion with endoglycosidase H, the majority of the wild-type E1 glycoprotein was endoglycosidase H sensitive, whereas the majority of the mutant E1 was processed to an endoglycosidase H-resistant, polylactosaminoglycan-containing form. The findings indicate that the wild-type E1 protein is specifically targeted to cis Golgi cisternae and are consistent with the assumption that the first membrane-spanning domain is required for targeting to the cis Golgi.
Machamer
C E
CE
Department of Pathology, Yale University School of Medicine, New Haven, CT 06510.
Mentone
S A
SA
Rose
J K
JK
Farquhar
M G
MG
D000818:Animals; D002460:Cell Line; D003332:Coronaviridae / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism / Q000648:ultrastructure*; D007124:Immunoenzyme Techniques; D001351:Infectious bronchitis virus / Q000235:genetics*; D008854:Microscopy, Electron; D009154:Mutation; D009844:Oligosaccharides / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D014616:Vaccinia virus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000032:analysis / Q000235:genetics*
D009844:Oligosaccharides; D014759:Viral Envelope Proteins