The effect of glucocorticoid on the subcellular localization, oligomerization, and processing of mouse mammary tumor virus envelope protein precursor Pr74.
Mouse lymphoma cell line W7MG1 is stably infected with mouse mammary tumor virus and produces the viral envelope glycoprotein precursor Pr74, but the mature envelope proteins gp52 and gp33, which are derived from Pr74 by posttranslational processing, are produced only when the cells are cultured with a glucocorticoid agonist. The current study demonstrated that even when W7MG1 cells are grown with hormone, the conversion of Pr74 to gp52 and gp33 is an inefficient process in this cell line. At least 2 h of exposure to glucocorticoid were required to induce the appearance of gp52 and gp33; furthermore, Pr74 labeled in the absence of hormone was not converted to gp52 and gp33 upon subsequent addition of hormone. RNA synthesis inhibitors blocked the hormonal induction of gp52 and gp33, indicating that the hormone acts by promoting the expression of a new gene(s) required for the production of gp52 and gp33, rather than by inhibiting the expression of a gene(s) that prevents processing of Pr74. Subcellular fractionation studies demonstrated that Pr74 produced in either the presence or absence of hormone was associated primarily with the ER, whereas gp52 and gp33 were found in the Golgi and plasma membrane fractions. The Pr74 molecules from W7MG1 cells grown either with or without glucocorticoid coimmunoprecipitated with BiP/GRP78 and sedimented as aggregates of heterogeneous size. In contrast, Pr74 from virus-producing GR3A mouse mammary tumor cells, which process Pr74 more efficiently, sedimented as apparent monomers, dimers, and trimers.(ABSTRACT TRUNCATED AT 250 WORDS)
Corey
J L
JL
Department of Pathology, University of Southern California Health Sciences Center, Los Angeles 90033.
Stallcup
M R
MR
D000818:Animals; D000957:Antigens, Viral, Tumor / Q000096:biosynthesis; D002352:Carrier Proteins / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D000091342:Endoplasmic Reticulum Chaperone BiP; D005938:Glucocorticoids / Q000502:physiology*; D006360:Heat-Shock Proteins; D008324:Mammary Tumor Virus, Mouse / Q000378:metabolism*; D051379:Mice; D018832:Molecular Chaperones; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000096:biosynthesis; D011965:Receptors, Glucocorticoid / Q000187:drug effects; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000378:metabolism*
D000957:Antigens, Viral, Tumor; D002352:Carrier Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; D005938:Glucocorticoids; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D018832:Molecular Chaperones; D011498:Protein Precursors; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D014759:Viral Envelope Proteins; C040612:glycoprotein 52 antigen, Mouse mammary tumor virus; D003609:Dactinomycin