Effects of inefficient cleavage of the signal sequence of HIV-1 gp 120 on its association with calnexin, folding, and intracellular transport.
The HIV-1 envelope glycoprotein gp120 displays inefficient intracellular transport, which is caused by its retention in the endoplasmic reticulum. Coexpression in insect cells (Sf9) of HIV-1 gp120 with calnexin has shown that their interaction was modulated by the signal sequence of HIV-1 gp120. gp120, with its natural signal sequence, showed a prolonged association with calnexin with a t1/2 of greater than 20 min. Replacement of the natural signal sequence with the signal sequence from mellitin led to a decreased time of association of gp120 with calnexin (t1/2 < 10 min). These different times of calnexin association coincided both with the folding of gp120 as measured by the ability of bind CD4 and with endoplasmic reticulum to Golgi transport as analyzed by the acquisition of partial endoglycosidase H resistance. Using a monospecific antibody to the HIV-1 gp120 natural signal peptide, we showed that calnexin associated with N-glycosylated but uncleaved gp120. Only after dissociation from calnexin was gp120 cleaved, but very inefficiently. Only the small proportion of signal-cleaved gp120 molecules acquired transport competence and were secreted. This is the first report demonstrating the effect of the signal sequence on calnexin association.
Li
Y
Y
Department of Zoology, Faculty of Science, University of Western Ontario, London, Canada.
Bergeron
J J
JJ
Luo
L
L
Ou
W J
WJ
Thomas
D Y
DY
Kang
C Y
CY
D000818:Animals; D001692:Biological Transport; D015704:CD4 Antigens / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism*; D037281:Calnexin; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006031:Glycosylation; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1 / Q000378:metabolism*; D006596:Hexosaminidases / Q000378:metabolism; D008555:Melitten / Q000378:metabolism; D017510:Protein Folding; D021382:Protein Sorting Signals / Q000378:metabolism*; D018411:Spodoptera
D015704:CD4 Antigens; D002135:Calcium-Binding Proteins; D015699:HIV Envelope Protein gp120; D021382:Protein Sorting Signals; D037281:Calnexin; D008555:Melitten; D006596:Hexosaminidases