The membrane glycoprotein G1 of Uukuniemi virus contains a signal for localization to the Golgi complex.
Members of the Bunyaviridae family acquire their envelopes by budding into the Golgi complex (GC). The accumulation of the membrane glycoproteins G1 and G2 in the GC probably determines the site of maturation. Here we have studied the intracellular transport and targeting to the GC of G1 and G2 of Uukuniemi virus, a member of the Phlebovirus genus, and report on their expression from cloned cDNAs either together or separately by using a T7 RNA polymerase-driven vaccinia virus expression system. When G1 and G2 were expressed together from a full-length cDNA as the p110 precursor, both proteins were localized to the Golgi complex, as evidenced by colocalization with the Golgi marker enzyme mannosidase II. Immunofluorescent staining indicated that G1 expressed alone also localized to the GC. However, pulse-chase experiments showed that G1 remained endoglycosidase H sensitive. G2 expressed alone remained associated with the endoplasmic reticulum (ER). G2 could be rescued from the ER and transported to the GC by coexpression with G1 from separate mRNAs. Coexpression also increased the efficiency of G1 transport to the GC. With none of the constructs could the glycoproteins be observed on the cell surface. These results show that efficient export of G1 and G2 from the ER requires coexpression of both proteins, in conformity with our previous results showing that G1 and G2 form heterodimeric complexes in the ER. Since G1 expressed alone is retained in the GC, we conclude that G1 contains a retention signal for localization to the GC. G2 might thus become associated with the GC indirectly via its interaction with G1.
Melin
L
L
Ludwig Institute for Cancer Research, Stockholm, Sweden.
Persson
R
R
Andersson
A
A
Bergström
A
A
Rönnholm
R
R
Pettersson
R F
RF
D001692:Biological Transport; D002460:Cell Line; D006056:Golgi Apparatus / Q000378:metabolism* / Q000821:virology; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids; D021382:Protein Sorting Signals / Q000378:metabolism*; D011995:Recombination, Genetic; D018066:Uukuniemi virus / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism*
D021382:Protein Sorting Signals; D014759:Viral Envelope Proteins