| The levels of ubiquitinated histone H2A are highly upregulated in transformed human cells: partial colocalization of uH2A clusters and PCNA/cyclin foci in a fraction of cells in S-phase. | |
| | |
MedLine Citation:
|
PMID: 7622605 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
|
The levels and distribution of ubiquitinated histone H2A (uH2A) have been studied in normal and transformed human cells using a monoclonal antibody (mAb E6C5) that reacts specifically with this ubiquitin conjugate as determined by two-dimensional gel western blotting and microsequencing. Immunoblotting experiments demonstrated that the levels of the protein are highly upregulated in SV40-transformed human fibroblasts (WI-38 SV40) and keratinocytes (K14) relative to their normal counterparts, a finding that was further confirmed by indirect immunofluorescence studies of formaldehyde/Triton X-100-treated cells, which showed that about 97% of the transformed cells and 26% of the normal populations reacted with the antibody to yield a fine granular nuclear staining associated with the chromatin. Transformed cells contained in addition clusters of uH2A that were quite abundant and that showed variable size, shape and distribution even within a single cell line. The clusters, which were rare in normal cells, did not colocalize with other known nuclear antigens and may correspond to novel nuclear domains where ubiquitination/deubiquitination takes place. Electron microscopic immunocytochemistry of K14 cells confirmed the existence of the clusters. Double immunofluorescence studies of K14 keratinocytes with proliferating cell nuclear antigen (PCNA)/cyclin antibodies, which react with the nuclei of cells engaged in DNA replication, showed partial colocalization of PCNA/cyclin foci and large uH2A clusters in about 14% of the S-phase cells, and these corresponded mainly to late S-phase cells. Inhibition of DNA replication with hydroxyurea resulted in an overall increase in the intensity of the uH2A staining as well as in a more clear colocalization of uH2A clusters and PCNA/cyclin foci. Taken together, the results support the contention that uH2A plays a role at some stage of DNA replication. |
| | |
Authors:
|
A P Vassilev; H H Rasmussen; E I Christensen; S Nielsen; J E Celis |
Related Documents
:
|
12566455 - Latrunculin b or atp depletion induces cofilin-dependent translocation of actin into nu... 3733875 - Dynamic behavior of histone h1 microinjected into hela cells. 3351925 - Newly synthesized small nuclear rnas appear transiently in the cytoplasm. 19244545 - Dependence on nuclear localization signals of the opioid growth factor receptor in the ... 9850925 - Pathological immuno-reactions of glial cells in alzheimer's disease and possible sites ... 23087405 - Availability of 25-hydroxyvitamin d(3) to apcs controls the balance between regulatory ... |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
|
Title: Journal of cell science Volume: 108 ( Pt 3) ISSN: 0021-9533 ISO Abbreviation: J. Cell. Sci. Publication Date: 1995 Mar |
Date Detail:
|
Created Date: 1995-08-29 Completed Date: 1995-08-29 Revised Date: 2006-11-15 |
Medline Journal Info:
|
Nlm Unique ID: 0052457 Medline TA: J Cell Sci Country: ENGLAND |
Other Details:
|
Languages: eng Pagination: 1205-15 Citation Subset: IM |
Affiliation:
|
Institute of Medical Biochemistry, University of Aarhus, Denmark. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
Amino Acid Sequence Antibodies, Monoclonal Antigens / genetics Cell Line Cell Line, Transformed Cell Transformation, Neoplastic DNA Replication Histones / genetics, immunology, metabolism* Humans Microscopy, Fluorescence Microscopy, Immunoelectron Molecular Sequence Data Proliferating Cell Nuclear Antigen / metabolism* S Phase Ubiquitins / genetics, immunology, metabolism* Up-Regulation |
| Chemical | |
Reg. No./Substance:
|
0/Antibodies, Monoclonal; 0/Antigens; 0/Histones; 0/Proliferating Cell Nuclear Antigen; 0/Ubiquitins; 0/chromatin conjugate protein A24 |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Suppression of a conditional mutation in alpha-tubulin by overexpression of two checkpoint genes.
Next Document: Regulation of Cdc2/cyclin B activation by Ran, a Ras-related GTPase.