1887

Abstract

We used a capture ELISA to quantify the E7 protein of human papillomavirus type 18 (HPV-18). In HeLa cells, which express low levels of immunoreactive E7 protein (iE7), iE7 had a mean half-life of 13·5 min. In HPV-18 E7 recombinant baculovirus (E7rec BV)-infected Sf21 cells, which express higher levels of E7, the half-life of iE7 was much longer (90 min and > 24 h, with two different E7rec BVs). For two transformed human cervical cell lines expressing HPV-18 E7, exposure of the cells to hydrocortisone resulted in a twofold increase in steady-state levels of the E7 protein: no similar effect was observed with progesterone, oestrogen or testosterone. The half-life of iE7 was unaltered by hydrocortisone or progesterone exposure. An immunoassay which distinguished Ser-phosphorylated E7 from E7 not phosphorylated at this residue (Serdephospho-E7), showed that in HeLa and Sf21 cells the majority of E7 was phosphorylated : the half-life of both species of E7 was similar in HeLa cells, but the half-life of Serdephospho-E7 was much shorter (90 min) in Sf21 cells than that of Serphospho-E7 (> 24 h). A HeLa- fibroblast fusion cell line with tumorigenic potential (CGL-1) had a similar ratio of dephospho-E7 to total E7 (0·06), as a similar fusion cell line (CGL-4) with no tumorigenic potential (0·03). We conclude that E7 is a labile phosphoprotein, and that the expression and steady-state level of the E7 protein in eukaryotic cells may be influenced by the hormonal environment of the cells.

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1994-07-01
2022-01-21
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