Arenavirus Stable Signal Peptide Is the Keystone Subunit for Glycoprotein Complex Organization

Bederka, Lydia H.; Bonhomme, Cyrille J.; Ling, Emily L.; Buchmeier, Michael J.

We assessed plasma membrane trafficking of the glycoprotein complex on live, transfected cells using flow cytometry (Fig. 3B ). The WT GPC, GP1/2 plus WT SSP, and HA SSP GPC samples supported the surface detection of the GP1 subunit. The GP1/2 and the GP1/2 plus SSP-HA samples resulted in mock levels of fluorescent intensity. Using the HA antibody to indirectly detect SSP, we detected wild-type GP1 levels of the HA signal with the HA SSP GPC sample, whereas the GP1/2 plus SSP-HA sample resulted in mock HA expression levels. Detection of the HA signal with the HA SSP GPC sample on live cells confirmed that our confocal microscopy results for SSP detection were not an artifact result from the fixation technique. (83.6) subunits. The LCMV SSP amino acid sequence, including the location of the inserted HA epitope, is shown. The "X" indicates where the membrane was sliced for antibody incubation and then reassembled for chemiluminescence processing and image acquisition.