Palmitylation of vesicular stomatitis virus G and Sindbis virus E1 glycoproteins has been studied in relation to the transport from the endoplasmic reticulum (ER) to the Golgi complex. Incubation of infected cells at 15 degrees C prevents the transport of newly synthesized membrane proteins from the ER to the Golgi (Saraste, J., and Kuismanen, E. (1984) Cell 38, 535-549). In these conditions, also palmitylation of G protein and of E1 glycoprotein is blocked. When the transport is restored by increasing the temperature, palmitylation occurs quickly and is followed by the complete trimming of peripheral mannose residues due to mannosidase I (a putative cis-Golgi function). Immunofluorescence analysis showed that the G glycoprotein accumulated at 15 degrees C in structures distinct from both ER and Golgi. These studies suggest that transport from the ER to the cis-Golgi involves intermediate compartments.