| Quantitative measurement of human papillomavirus type 16 e5 oncoprotein levels in epithelial cell lines by mass spectrometry. | |
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MedLine Citation:
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PMID: 22740411 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The high-risk human papillomavirus type 16 (HPV-16) E5 protein (16E5) induces tumors in a transgenic mouse model and may contribute to early stages of cervical carcinogenesis. Although high-risk E5 expression is generally thought to be lost during the progression to cervical carcinoma following integration of HPV DNA into the host genome, episomal viral DNA has been documented in a subset of HPV-16-positive malignant lesions. Numerous studies have shown that transcripts that could potentially encode 16E5 are present in cervical biopsy specimens and cervical cancer cell lines, but the presence of E5 protein has been demonstrated in only two reports that have not been corroborated. In the present study, we show that trypsin cleavage of 16E5 generates a unique four-amino-acid C-terminal peptide (FLIT) that serves as a marker for E5 expression in transfected cells and epithelial cell lines containing integrated and episomal HPV-16 DNA. Following trypsin cleavage, reversed-phase chromatography and mass spectrometry (MS) were used to detect FLIT. Immunoprecipitation assays using a newly generated anti-16E5 antibody confirmed that 16E5 was solely responsible for the FLIT signal, and deuterated FLIT peptide provided an internal standard that enabled us to quantify the number of 16E5 molecules per cell. We show that 16E5 is expressed in the Caski but not in the SiHa cervical cancer cell line, suggesting that 16E5 may contribute to the malignant phenotype of some cervical cancers, even in cells exclusively containing an integrated HPV genome. |
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Authors:
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Ziad Sahab; Sawali R Sudarshan; Xuefeng Liu; YiYu Zhang; Alexander Kirilyuk; Christopher M Kamonjoh; Vera Simic; Yuhai Dai; Stephen W Byers; John Doorbar; Frank A Suprynowicz; Richard Schlegel |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2012-06-27 |
Journal Detail:
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Title: Journal of virology Volume: 86 ISSN: 1098-5514 ISO Abbreviation: J. Virol. Publication Date: 2012 Sep |
Date Detail:
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Created Date: 2012-08-10 Completed Date: 2012-11-05 Revised Date: 2013-04-15 |
Medline Journal Info:
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Nlm Unique ID: 0113724 Medline TA: J Virol Country: United States |
Other Details:
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Languages: eng Pagination: 9465-73 Citation Subset: IM |
Affiliation:
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Department of Pathology, Georgetown University Medical School, Washington, DC, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Animals Cell Line, Transformed Cell Line, Tumor Epithelial Cells / chemistry*, metabolism Female Human papillomavirus 16 / chemistry*, genetics, metabolism Humans Mass Spectrometry / methods Mice Molecular Sequence Data Oncogene Proteins, Viral / analysis*, genetics, metabolism Peptide Mapping Uterine Cervical Neoplasms / chemistry, genetics, metabolism, virology |
| Grant Support | |
ID/Acronym/Agency:
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R01-CA053371/CA/NCI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Oncogene Proteins, Viral; 0/oncogene protein E5, Human papillomavirus type 16 |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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