RETENTION OF A CIS GOLGI PROTEIN REQUIRES POLAR RESIDUES ON ONE FACE OF A PREDICTED ALPHA-HELIX IN THE TRANSMEMBRANE DOMAIN

The first membrane-spanning domain (ml) of the model cis Golgi protein M (formerly called El) from the avian coronavirus infectious bronchitis virus is required for targeting to the Golgi complex. When inserted in place of the membrane-spanning domain of a plasma membrane protein (vesicular stomatitis virus G protein), the chimeric protein (''Gm1'') is retained in the Golgi complex of transfected cells. To determine the precise features of the ml domain responsible for Golgi targeting, we produced single amino acid substitutions in ml and analyzed their effects on localization of Gm1. Expression at the plasma membrane was used as the criterion for loss of Golgi retention. Rates of oligosaccharide processing were used as a measure of rate and efficiency of transport through the Golgi complex. We identified four uncharged polar residues that are critical for Golgi retention of Gm1 (Asn465, Thr469, Thr476, and Gln480). These residues line one face of a predicted alpha-helix. Interestingly, when the ml domain of the homologous M protein from mouse hepatitis virus is inserted into the G protein reporter, the chimeric protein is not efficiently retained in the Golgi complex, but transported to the cell surface. Although it possesses three of the four residues we identified as important in the avian ml sequence, other residues in the membrane-spanning domain from the mouse protein must prevent efficient recognition of the polar face within the lipid bilayer of the cis Golgi.