Document Detail


Bimolecular fluorescence complementation analysis of eukaryotic fusion products.
MedLine Citation:
PMID:  20590528     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND INFORMATION: Cell fusion is known to underlie key developmental processes in humans and is postulated to contribute to tissue maintenance and even carcinogenesis. The mechanistic details of cell fusion, especially between different cell types, have been difficult to characterize because of the dynamic nature of the process and inadequate means to track fusion products over time. Here we introduce an inducible system for detecting and tracking live cell fusion products in vitro and potentially in vivo. This system is based on BiFC (bimolecular fluorescence complementation) analysis. In this approach, two proteins that can interact with each other are joined to fragments of a fluorescent protein and are expressed in separate cells. The interaction of said proteins after cell fusion produces a fluorescent signal, enabling the identification and tracking of fusion products over time.
RESULTS: Long-term tracking of fused p53-deficient cells revealed that hybrid cells were capable of proliferation. In some cases, proliferation was preceded by nuclear fusion and division was asymmetric (69%+/-2% of proliferating hybrids), suggesting chromosomal instability. In addition, asymmetric division following proliferation could give rise to progeny indistinguishable from unfused counterparts.
CONCLUSIONS: These results support the possibility that the chromosomal instability characteristic of tumour cells may be incurred as a consequence of cell fusion and suggest that the role of cell fusion in carcinogenesis may have been masked to this point for lack of an inducible method to track cell fusion. In sum, the BiFC-based approach described here allows for comprehensive studies of the mechanism and biological impact of cell fusion in nature.
Authors:
Ho-Pi Lin; Claudius Vincenz; Kevin W Eliceiri; Tom K Kerppola; Brenda M Ogle
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-08-06
Journal Detail:
Title:  Biology of the cell / under the auspices of the European Cell Biology Organization     Volume:  102     ISSN:  1768-322X     ISO Abbreviation:  Biol. Cell     Publication Date:  2010 Sep 
Date Detail:
Created Date:  2010-08-09     Completed Date:  2010-11-01     Revised Date:  2011-08-03    
Medline Journal Info:
Nlm Unique ID:  8108529     Medline TA:  Biol Cell     Country:  England    
Other Details:
Languages:  eng     Pagination:  525-37     Citation Subset:  IM    
Affiliation:
Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI 53706, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
COS Cells
Cell Division
Cell Fusion*
Cell Proliferation
Cercopithecus aethiops
Chromosomal Instability*
Fluorescence
Hybrid Cells / cytology,  physiology*
Luminescent Measurements / methods*
Luminescent Proteins / metabolism
Microscopy, Fluorescence / methods*
Protein Binding
Recombinant Fusion Proteins / metabolism
Grant Support
ID/Acronym/Agency:
AI057358/AI/NIAID NIH HHS; HL089679/HL/NHLBI NIH HHS; R01 GM086213/GM/NIGMS NIH HHS; R01 GM086213-04/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute
Chemical
Reg. No./Substance:
0/Luminescent Proteins; 0/Recombinant Fusion Proteins
Comments/Corrections

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