Characterization of the biosynthesis of human immunodeficiency virus type 1 Env from infected T-cells and the effects of glucose trimming of Env on virion infectivity.
HIV (human immunodeficiency virus)-1 Env is displayed on the surface of infected cells and subsequently incorporated into virions, which is necessary for the initiation of a viral infection by recognition of the CD4 and the chemokine receptors (such as CCR5 or CXCR4) on the surface of new target cells. As a type 1 integral membrane glycoprotein, Env is cotranslationally translocated into the endoplasmic reticulum. In this report, we characterized the synthesis of Env, which did not occur at a constant rate but by translational/translocational pausing that has not previously been shown with a viral encoded glycoprotein. Overall translation was not impeded by the presence of the reducing agent dithiothreitol in vivo, although this did influence the cleavage of the precursor gp160 into its mature form, gp120. Env interacts transiently with resident components of the endoplasmic reticulum such as calnexin, which had maximal association at a 10-min post-translation. Addition of the glucosidase inhibitor, castanospermine, failed to significantly influence the association of Env with calnexin, consistent with the notion that calnexin recognizes components other than alpha-terminal glucose. Moreover, castanospermine treatment failed to affect the infectivity of virions. Taken together, this report demonstrates the existence of translational/translocational pausing for a viral glycoprotein and suggests that trimming of glucose from HIV-1 Env is not essential for the initiation of virus infection.
Dettenhofer
M
M
Department of Molecular Microbiology and Immunology, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205, USA.
Yu
X F
XF
D000255:Adenosine Triphosphate / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D004220:Disulfides / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology*; D006031:Glycosylation; D019302:HIV Envelope Protein gp160 / Q000096:biosynthesis*; D015497:HIV-1 / Q000472:pathogenicity*; D007212:Indolizines / Q000494:pharmacology; D011485:Protein Binding; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D021381:Protein Transport / Q000187:drug effects*; D013601:T-Lymphocytes / Q000821:virology*; D014771:Virion / Q000472:pathogenicity; D054836:Indolizidines
D000255:Adenosine Triphosphate; D002135:Calcium-Binding Proteins; D037281:Calnexin; D004220:Disulfides; D004229:Dithiothreitol; D005947:Glucose; D019302:HIV Envelope Protein gp160; D007212:Indolizines; C037806:castanospermine; D054836:Indolizidines