Analysis of the hemagglutinin glycoprotein from mutants of vaccinia virus that accumulates on the nuclear envelope.

We isolated mutants whose vaccinia hemagglutinin (HA) accumulates on nuclear envelopes and the rough endoplasmic reticulum. Mutant HA must be blocked at a pre-Golgi step because it has high-mannose-type carbohydrates but no fucose. Neither N- nor O-glycosidically linked carbohydrates are involved in the transport defect of the mutant HA, because tunicamycin, an inhibitor of N-type glycosylation, has no effect, and O-type glycosylation takes place in the Golgi organelle. The unglycosylated form of the mutant HA synthesized in the presence of tunicamycin is 3000 daltons larger than the wild type. This higher molecular weight is related to the transport defect. HAs translated in vitro also show this difference, evidence that it reflects mutation in the HA structural gene. Portions of HAs that project into the cytoplasm seem to account for this weight difference. Thus the cytoplasmic tail of glycoprotein has an important function in transport out of the rough endoplasmic reticulum.