A monoclonal antibody against p53 cross-reacts with processing bodies

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The p53 tumor suppressor protein is an important regulator of cell proliferation and apoptosis. p53 can be found in the nucleus and in the cytosol, and the subcellular location is key to control p53 function. In this work, we found that a widely used monoclonal antibody against p53, termed Pab 1801...

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Autores principales:	Thomas, M.G., Luchelli, L., Pascual, M., Gottifredi, V., Boccaccio, G.L.
Formato:	Artículo publishedVersion
Lenguaje:	Inglés
Publicado:	2012
Materias:	cell marker cycloheximide decapping enzyme 1a decapping enzyme 1b decapping enzyme 2 epitope exoribonuclease exoribonuclease 1 messenger RNA monoclonal antibody pantropic antibody 1801 protein 4ET protein Hedls protein p53 protein p54 small interfering RNA unclassified drug TP53 protein, human animal cell animal tissue article cell component cell disruption cell line cell stimulation cell strain HCT116 cellular distribution concentration (parameters) controlled study cross reaction dissolution Drosophila embryo fetus gene control gene silencing genetic transfection human human cell image analysis immunohistochemistry intracellular signaling nonhuman polysome processing body protein analysis protein localization rat upregulation animal antibody specificity chemistry cytoplasm immunology metabolism Sprague Dawley rat tumor cell line Rattus Rodentia Animals Antibodies, Monoclonal, Murine-Derived Antibody Specificity Cell Line, Tumor Cytoplasm Epitopes Humans Immunohistochemistry Rats Rats, Sprague-Dawley Tumor Suppressor Protein p53
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_19326203_v7_n5_p_Thomas
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paperaa:paper_19326203_v7_n5_p_Thomas
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spelling	paperaa:paper_19326203_v7_n5_p_Thomas2023-06-12T16:51:39Z A monoclonal antibody against p53 cross-reacts with processing bodies PLoS ONE 2012;7(5) Thomas, M.G. Luchelli, L. Pascual, M. Gottifredi, V. Boccaccio, G.L. cell marker cycloheximide decapping enzyme 1a decapping enzyme 1b decapping enzyme 2 epitope exoribonuclease exoribonuclease 1 messenger RNA monoclonal antibody pantropic antibody 1801 protein 4ET protein Hedls protein p53 protein p54 small interfering RNA unclassified drug epitope protein p53 TP53 protein, human animal cell animal tissue article cell component cell disruption cell line cell stimulation cell strain HCT116 cellular distribution concentration (parameters) controlled study cross reaction dissolution Drosophila embryo fetus gene control gene silencing genetic transfection human human cell image analysis immunohistochemistry intracellular signaling nonhuman polysome processing body protein analysis protein localization rat upregulation animal antibody specificity chemistry cytoplasm immunology metabolism Sprague Dawley rat tumor cell line Rattus Rodentia Animals Antibodies, Monoclonal, Murine-Derived Antibody Specificity Cell Line, Tumor Cytoplasm Epitopes Humans Immunohistochemistry Rats Rats, Sprague-Dawley Tumor Suppressor Protein p53 The p53 tumor suppressor protein is an important regulator of cell proliferation and apoptosis. p53 can be found in the nucleus and in the cytosol, and the subcellular location is key to control p53 function. In this work, we found that a widely used monoclonal antibody against p53, termed Pab 1801 (Pan antibody 1801) yields a remarkable punctate signal in the cytoplasm of several cell lines of human origin. Surprisingly, these puncta were also observed in two independent p53-null cell lines. Moreover, the foci stained with the Pab 1801 were present in rat cells, although Pab 1801 recognizes an epitope that is not conserved in rodent p53. In contrast, the Pab 1801 nuclear staining corresponded to genuine p53, as it was upregulated by p53-stimulating drugs and absent in p53-null cells. We identified the Pab 1801 cytoplasmic puncta as P Bodies (PBs), which are involved in mRNA regulation. We found that, in several cell lines, including U2OS, WI38, SK-N-SH and HCT116, the Pab 1801 puncta strictly colocalize with PBs identified with specific antibodies against the PB components Hedls, Dcp1a, Xrn1 or Rck/p54. PBs are highly dynamic and accordingly, the Pab 1801 puncta vanished when PBs dissolved upon treatment with cycloheximide, a drug that causes polysome stabilization and PB disruption. In addition, the knockdown of specific PB components that affect PB integrity simultaneously caused PB dissolution and the disappearance of the Pab 1801 puncta. Our results reveal a strong cross-reactivity of the Pab 1801 with unknown PB component(s). This was observed upon distinct immunostaining protocols, thus meaning a major limitation on the use of this antibody for p53 imaging in the cytoplasm of most cell types of human or rodent origin. © 2012 Thomas et al. Fil:Thomas, M.G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Luchelli, L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Boccaccio, G.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2012 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_19326203_v7_n5_p_Thomas
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language	Inglés
orig_language_str_mv	eng
topic	cell marker cycloheximide decapping enzyme 1a decapping enzyme 1b decapping enzyme 2 epitope exoribonuclease exoribonuclease 1 messenger RNA monoclonal antibody pantropic antibody 1801 protein 4ET protein Hedls protein p53 protein p54 small interfering RNA unclassified drug epitope protein p53 TP53 protein, human animal cell animal tissue article cell component cell disruption cell line cell stimulation cell strain HCT116 cellular distribution concentration (parameters) controlled study cross reaction dissolution Drosophila embryo fetus gene control gene silencing genetic transfection human human cell image analysis immunohistochemistry intracellular signaling nonhuman polysome processing body protein analysis protein localization rat upregulation animal antibody specificity chemistry cytoplasm immunology metabolism Sprague Dawley rat tumor cell line Rattus Rodentia Animals Antibodies, Monoclonal, Murine-Derived Antibody Specificity Cell Line, Tumor Cytoplasm Epitopes Humans Immunohistochemistry Rats Rats, Sprague-Dawley Tumor Suppressor Protein p53
spellingShingle	cell marker cycloheximide decapping enzyme 1a decapping enzyme 1b decapping enzyme 2 epitope exoribonuclease exoribonuclease 1 messenger RNA monoclonal antibody pantropic antibody 1801 protein 4ET protein Hedls protein p53 protein p54 small interfering RNA unclassified drug epitope protein p53 TP53 protein, human animal cell animal tissue article cell component cell disruption cell line cell stimulation cell strain HCT116 cellular distribution concentration (parameters) controlled study cross reaction dissolution Drosophila embryo fetus gene control gene silencing genetic transfection human human cell image analysis immunohistochemistry intracellular signaling nonhuman polysome processing body protein analysis protein localization rat upregulation animal antibody specificity chemistry cytoplasm immunology metabolism Sprague Dawley rat tumor cell line Rattus Rodentia Animals Antibodies, Monoclonal, Murine-Derived Antibody Specificity Cell Line, Tumor Cytoplasm Epitopes Humans Immunohistochemistry Rats Rats, Sprague-Dawley Tumor Suppressor Protein p53 Thomas, M.G. Luchelli, L. Pascual, M. Gottifredi, V. Boccaccio, G.L. A monoclonal antibody against p53 cross-reacts with processing bodies
topic_facet	cell marker cycloheximide decapping enzyme 1a decapping enzyme 1b decapping enzyme 2 epitope exoribonuclease exoribonuclease 1 messenger RNA monoclonal antibody pantropic antibody 1801 protein 4ET protein Hedls protein p53 protein p54 small interfering RNA unclassified drug epitope protein p53 TP53 protein, human animal cell animal tissue article cell component cell disruption cell line cell stimulation cell strain HCT116 cellular distribution concentration (parameters) controlled study cross reaction dissolution Drosophila embryo fetus gene control gene silencing genetic transfection human human cell image analysis immunohistochemistry intracellular signaling nonhuman polysome processing body protein analysis protein localization rat upregulation animal antibody specificity chemistry cytoplasm immunology metabolism Sprague Dawley rat tumor cell line Rattus Rodentia Animals Antibodies, Monoclonal, Murine-Derived Antibody Specificity Cell Line, Tumor Cytoplasm Epitopes Humans Immunohistochemistry Rats Rats, Sprague-Dawley Tumor Suppressor Protein p53
description	The p53 tumor suppressor protein is an important regulator of cell proliferation and apoptosis. p53 can be found in the nucleus and in the cytosol, and the subcellular location is key to control p53 function. In this work, we found that a widely used monoclonal antibody against p53, termed Pab 1801 (Pan antibody 1801) yields a remarkable punctate signal in the cytoplasm of several cell lines of human origin. Surprisingly, these puncta were also observed in two independent p53-null cell lines. Moreover, the foci stained with the Pab 1801 were present in rat cells, although Pab 1801 recognizes an epitope that is not conserved in rodent p53. In contrast, the Pab 1801 nuclear staining corresponded to genuine p53, as it was upregulated by p53-stimulating drugs and absent in p53-null cells. We identified the Pab 1801 cytoplasmic puncta as P Bodies (PBs), which are involved in mRNA regulation. We found that, in several cell lines, including U2OS, WI38, SK-N-SH and HCT116, the Pab 1801 puncta strictly colocalize with PBs identified with specific antibodies against the PB components Hedls, Dcp1a, Xrn1 or Rck/p54. PBs are highly dynamic and accordingly, the Pab 1801 puncta vanished when PBs dissolved upon treatment with cycloheximide, a drug that causes polysome stabilization and PB disruption. In addition, the knockdown of specific PB components that affect PB integrity simultaneously caused PB dissolution and the disappearance of the Pab 1801 puncta. Our results reveal a strong cross-reactivity of the Pab 1801 with unknown PB component(s). This was observed upon distinct immunostaining protocols, thus meaning a major limitation on the use of this antibody for p53 imaging in the cytoplasm of most cell types of human or rodent origin. © 2012 Thomas et al.
format	Artículo Artículo publishedVersion
author	Thomas, M.G. Luchelli, L. Pascual, M. Gottifredi, V. Boccaccio, G.L.
author_facet	Thomas, M.G. Luchelli, L. Pascual, M. Gottifredi, V. Boccaccio, G.L.
author_sort	Thomas, M.G.
title	A monoclonal antibody against p53 cross-reacts with processing bodies
title_short	A monoclonal antibody against p53 cross-reacts with processing bodies
title_full	A monoclonal antibody against p53 cross-reacts with processing bodies
title_fullStr	A monoclonal antibody against p53 cross-reacts with processing bodies
title_full_unstemmed	A monoclonal antibody against p53 cross-reacts with processing bodies
title_sort	monoclonal antibody against p53 cross-reacts with processing bodies
publishDate	2012
url	http://hdl.handle.net/20.500.12110/paper_19326203_v7_n5_p_Thomas
work_keys_str_mv	AT thomasmg amonoclonalantibodyagainstp53crossreactswithprocessingbodies AT luchellil amonoclonalantibodyagainstp53crossreactswithprocessingbodies AT pascualm amonoclonalantibodyagainstp53crossreactswithprocessingbodies AT gottifrediv amonoclonalantibodyagainstp53crossreactswithprocessingbodies AT boccacciogl amonoclonalantibodyagainstp53crossreactswithprocessingbodies AT thomasmg monoclonalantibodyagainstp53crossreactswithprocessingbodies AT luchellil monoclonalantibodyagainstp53crossreactswithprocessingbodies AT pascualm monoclonalantibodyagainstp53crossreactswithprocessingbodies AT gottifrediv monoclonalantibodyagainstp53crossreactswithprocessingbodies AT boccacciogl monoclonalantibodyagainstp53crossreactswithprocessingbodies
_version_	1769810355082493952

A monoclonal antibody against p53 cross-reacts with processing bodies

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