Abstract:
:Transfection of the E6 and E7 genes of the high-risk human papillomaviruses (HPVs) into primary genital keratinocytes generates colonies of proliferating cells which are resistant to calcium- and serum-induced terminal differentiation. To genetically map the HPV gene(s) responsible for this cellular phenotype, we cloned cDNAs for full-length E6, full-length E7, four truncated E6 splice variants (E6I to E6IV), and a series of E6 C-terminal truncation mutants into a simian virus 40 expression vector. The E6 proteins were tagged with the AU1 epitope at the C terminus to verify their expression in COS cells by immunoprecipitation and immunofluorescence. Transfection of the full-length E6 protein, either wild type or epitope tagged, induced calcium- and serum-resistant keratinocyte colonies, but the truncated E6 variants and full-length E7 protein did not. E6-induced colonies, while altered in response to serum and calcium, could not be established into cell lines without the combined presence of the E7 protein, further exemplifying the independent roles of E6 and E7 in cell differentiation and cell proliferation. The E6 C-terminal deletion mutants defined two distinct functional domains between amino acids 120 and 151. Amino acids 120 to 151 contained an apparent bipartite nuclear localization signal, whereas amino acids 132 to 141 were required for the induction of resistance to calcium- and serum-induced differentiation and for immortalization of human keratinocytes in conjunction with E7.
journal_name
J Viroljournal_title
Journal of virologyauthors
Sherman L,Schlegel Rdoi
10.1128/JVI.70.5.3269-3279.1996subject
Has Abstractpub_date
1996-05-01 00:00:00pages
3269-79issue
5eissn
0022-538Xissn
1098-5514journal_volume
70pub_type
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pub_type: 杂志文章
doi:10.1128/JVI.01971-14
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journal_title:Journal of virology
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doi:10.1128/JVI.02897-14
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.5.4157-4160.1997
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pub_type: 杂志文章
doi:10.1128/JVI.79.5.3187-3194.2005
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journal_title:Journal of virology
pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.75.5.2462-2467.2001
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journal_title:Journal of virology
pub_type: 杂志文章
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journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:2015-05-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.75.9.4453-4458.2001
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journal_title:Journal of virology
pub_type: 杂志文章
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更新日期:2014-03-01 00:00:00
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journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00777-13
更新日期:2013-08-01 00:00:00