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PMID: 1662388 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Human papillomavirus 16 E7 protein is associated with the nuclear matrix.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 24 ·1991-12-15 ·Pages 11217-21

Greenfield I, Nickerson J, Penman S, Stanley M

Abstract

The cellular localization of the human papillomavirus (HPV)-16 E7 gene product in the cell lines CaSki and SiHa has been determined by both biochemical and immunocytochemical methods. These measurements show E7 to be localized in the cell nucleus, specifically with the nonchromatin nuclear structure or nuclear matrix. This localization of E7 required an unambiguous fractionation of the nuclear constituents. This was achieved by using a gentle sequential fractionation procedure to prepare the scaffold consisting of the nuclear matrix and intermediate filaments (NM-IF). Chromatin was cleaved with nuclease and the resulting nucleosomes eluted with 0.25 M ammonium sulfate. Immunostaining of cells after this extraction procedure with monoclonal antibodies (mAbs) to E7 revealed a fine grained, punctate nuclear fluorescence in CaSki and SiHa, which was absent in normal cervical keratinocytes and the HPV-negative cell line C33.1. Western blots of cell fractions with these mAbs showed that E7 was localized in the NM-IF fraction in SiHa and CaSki but was not detected in HPV-negative cells. A second protein of slightly higher mobility is identified by these antisera in HPV-16-containing cells. The data suggest that the previous inability to directly visualize E7 by immunocytology is due to the masking of epitopes by cellular components and not to low levels of protein.

MeSH Terms
Blotting, Western Cell Line Fluorescent Antibody Technique Humans Immunohistochemistry Molecular Weight Nuclear Matrix/physiology,ultrastructure Oncogene Proteins, Viral/analysis,metabolism Papillomaviridae/genetics Papillomavirus E7 Proteins
Chemicals
Oncogene Proteins, Viral Papillomavirus E7 Proteins oncogene protein E7, Human papillomavirus type 16
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Greenfield I
Department of Pathology, University of Cambridge, United Kingdom.
Nickerson J
Penman S
Stanley M
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.24.11217
Published
1991-12-15
Pages
11217-21
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC53105
Subset
IM
Grants
NCI NIH HHS · CA08416 · United States
NCI NIH HHS · CA45480 · United States
External Links
PubMed source DOI: 10.1073/pnas.88.24.11217 PMC Full Text View journal details
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