Bimolecular fluorescence complementation analysis of eukaryotic fusion products

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.

[1]  J. Platt,et al.  Biological implications of cell fusion , 2005, Nature Reviews Molecular Cell Biology.

[2]  T. Warner.  Cell hybridizaiton: an explanation for the phenotypic diversity of certain tumours. , 1975, Medical hypotheses.

[3]  Y. Gluzman.  SV40-transformed simian cells support the replication of early SV40 mutants , 1981, Cell.

[4]  Y. Lazebnik,et al.  Cell-to-cell fusion as a link between viruses and cancer , 2007, Nature Reviews Cancer.

[5]  Alan Shiels,et al.  The stratified syncytium of the vertebrate lens , 2009, Journal of Cell Science.

[6]  M. Harada,et al.  Purified human hematopoietic stem cells contribute to the generation of cardiomyocytes through cell fusion , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[7]  B. Fleischmann,et al.  Bone marrow–derived hematopoietic cells generate cardiomyocytes at a low frequency through cell fusion, but not transdifferentiation , 2004, Nature Medicine.

[8]  J. Bourhis,et al.  Enhancement of radiation response in p53-deficient cancer cells by the Aurora-B kinase inhibitor AZD1152 , 2007, Oncogene.

[9]  H. Blau,et al.  Gene expression and cell fusion analyzed by lacZ complementation in mammalian cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Jesse J. Lipp,et al.  Histone H3 serine 10 phosphorylation by Aurora B causes HP1 dissociation from heterochromatin , 2005, Nature.

[11]  K. Itoh,et al.  Bach proteins belong to a novel family of BTB-basic leucine zipper transcription factors that interact with MafK and regulate transcription through the NF-E2 site , 1996, Molecular and cellular biology.

[12]  K. Benirschke,et al.  Pathology of the Human Placenta , 1974, Springer New York.

[13]  A. Báez-Saldaña,et al.  Human immunodeficiency virus envelope-dependent cell-cell fusion: a quantitative fluorescence cytometric assay. , 2002, Cytometry.

[14]  K. Abromeit.  Music Received , 2023, Notes.

[15]  G. Vassilopoulos,et al.  Transplanted bone marrow regenerates liver by cell fusion , 2003, Nature.

[16]  T. Kerppola,et al.  Visualization of molecular interactions by fluorescence complementation , 2006, Nature Reviews Molecular Cell Biology.

[17]  C. Murry,et al.  Myogenic fusion of human bone marrow stromal cells, but not hematopoietic cells. , 2004, Blood.

[18]  I. Sargent,et al.  Trophoblast deportation in human pregnancy--its relevance for pre-eclampsia. , 1999, Placenta.

[19]  Keiichiro Yoshida,et al.  TPA-induced multinucleation of a mesenchymal stem cell-like clone is mediated primarily by karyokinesis without cytokinesis, although cell-cell fusion also occurs. , 2007, European journal of cell biology.

[20]  Chang‐Deng Hu,et al.  Visualization of interactions among bZIP and Rel family proteins in living cells using bimolecular fluorescence complementation. , 2002, Molecular cell.

[21]  M. Grompe,et al.  Ploidy Reductions in Murine Fusion-Derived Hepatocytes , 2009, PLoS genetics.

[22]  G. Parris.  Clinically significant cancer evolves from transient mutated and/or aneuploid neoplasia by cell fusion to form unstable syncytia that give rise to ecologically viable parasite species. , 2005, Medical hypotheses.

[23]  A. Verreault,et al.  De novo nucleosome assembly: new pieces in an old puzzle. , 2000, Genes & development.

[24]  Elizabeth H. Chen,et al.  Unveiling the Mechanisms of Cell-Cell Fusion , 2005, Science.

[25]  I. Sargent,et al.  Placental debris, oxidative stress and pre-eclampsia. , 2000, Placenta.

[26]  J. Platt,et al.  Spontaneous fusion of cells between species yields transdifferentiation and retroviral transfer in vivo , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  Klaus Pfeffer,et al.  Fusion of bone-marrow-derived cells with Purkinje neurons, cardiomyocytes and hepatocytes , 2003, Nature.

[28]  M. Grompe,et al.  Cell fusion is the principal source of bone-marrow-derived hepatocytes , 2003, Nature.

[29]  K. Ghosh,et al.  Reproducible methodology for the isolation of mesenchymal stem cells from human umbilical cord and its potential for cardiomyocyte generation , 2008, Journal of tissue engineering and regenerative medicine.

[30]  H. Blau,et al.  Extensive fusion of haematopoietic cells with Purkinje neurons in response to chronic inflammation , 2008, Nature Cell Biology.

[31]  Qinghua Shi,et al.  Chromosome nondisjunction yields tetraploid rather than aneuploid cells in human cell lines , 2005, Nature.

[32]  Robin C. Allshire,et al.  Dimerisation of a chromo shadow domain and distinctions from the chromodomain as revealed by structural analysis , 2000, Current Biology.

[33]  T. Kerppola,et al.  Changes in the Distributions and Dynamics of Polycomb Repressive Complexes during Embryonic Stem Cell Differentiation , 2008, Molecular and Cellular Biology.

[34]  Helen M. Blau,et al.  Cytoplasmic activation of human nuclear genes in stable heterocaryons , 1983, Cell.

[35]  M. Boothby,et al.  Cytological localization of chorionic gonadotropin alpha and placental lactogen mRNAs during development of the human placenta , 1982, The Journal of cell biology.

[36]  J. Green,et al.  A single vector containing modified cre recombinase and LOX recombination sequences for inducible tissue-specific amplification of gene expression. , 2001, Nucleic acids research.

[37]  T. Kerppola,et al.  Different polycomb group CBX family proteins associate with distinct regions of chromatin using nonhomologous protein sequences , 2008, Proceedings of the National Academy of Sciences.

[38]  F. Camargo,et al.  Hematopoietic myelomonocytic cells are the major source of hepatocyte fusion partners. , 2004, The Journal of clinical investigation.

[39]  C. Landry,et al.  An in Vivo Map of the Yeast Protein Interactome , 2008, Science.

[40]  M. Kalējs,et al.  Activation of meiosis-specific genes is associated with depolyploidization of human tumor cells following radiation-induced mitotic catastrophe. , 2009, Cancer research.