Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein

Autores
Ahn, Jinwoo; Poyurowsky, Masha V.; Baptiste, Nicole; Beckerman, Rachel; Cain, Christine; Mattia, Melissa; McKinney, Kristine; Zhou, Jianmin; Zupnick, Andrew; Gottifredi, Vanesa; Prives, Carol
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Both sequence-specific DNA binding and exonuclease activities have been mapped to the central conserved core domain of p53. To gain more information about these two activities a series of mutants were generated that changed core domain histidine residues. Of these mutants, only one, H115N p53, showed markedly reduced exonuclease activity (ca. 15% of wild-type). Surprisingly, purified H115N p53 protein was found to be significantly more potent than wild-type p53 in binding to DNA by several criteria including gel mobility shift assay, filter binding and DNase I footprinting. Interestingly as well, non-specific DNA binding by the core domain of H115N p53 is superior to that of wild-type p53. To study H115N p53 in vivo, clones of H1299 cells expressing tetracycline regulated wild-type or H115N p53 were generated. H115N was both more potent than wild-type p53 in inducing p53 target genes such as p21 and PIG3 and was also more effective in arresting cells in G1. Unexpectedly, in contrast to wild-type p53, H115N p53 was markedly impaired in causing apoptosis when cells were subjected to DNA damage. Our results indicate that the exonuclease activity and transcriptional activation functions of p53 can be separated. They also extend previous findings showing that cell cycle arrest and apoptosis are separable functions of p53. Finally, these experiments confirm that DNA binding and xonuclease activities are distinct features of the p53 core domain.
Fil: Ahn, Jinwoo. University Of Pittsburgh; Estados Unidos
Fil: Poyurowsky, Masha V.. Columbia University; Estados Unidos
Fil: Baptiste, Nicole. Columbia University; Estados Unidos
Fil: Beckerman, Rachel. Columbia University; Estados Unidos
Fil: Cain, Christine. Columbia University; Estados Unidos
Fil: Mattia, Melissa. Mount Sinai School of Medicine; Estados Unidos
Fil: McKinney, Kristine. Harvard Medical School; Estados Unidos
Fil: Zhou, Jianmin. Columbia University; Estados Unidos
Fil: Zupnick, Andrew. Columbia University; Estados Unidos
Fil: Gottifredi, Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Prives, Carol. Columbia University; Estados Unidos
Materia
P53
Apoptosis
Exonuclease Activity
Dna Binding
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/20640
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/20640
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 proteinAhn, JinwooPoyurowsky, Masha V.Baptiste, NicoleBeckerman, RachelCain, ChristineMattia, MelissaMcKinney, KristineZhou, JianminZupnick, AndrewGottifredi, VanesaPrives, CarolP53ApoptosisExonuclease ActivityDna Bindinghttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Both sequence-specific DNA binding and exonuclease activities have been mapped to the central conserved core domain of p53. To gain more information about these two activities a series of mutants were generated that changed core domain histidine residues. Of these mutants, only one, H115N p53, showed markedly reduced exonuclease activity (ca. 15% of wild-type). Surprisingly, purified H115N p53 protein was found to be significantly more potent than wild-type p53 in binding to DNA by several criteria including gel mobility shift assay, filter binding and DNase I footprinting. Interestingly as well, non-specific DNA binding by the core domain of H115N p53 is superior to that of wild-type p53. To study H115N p53 in vivo, clones of H1299 cells expressing tetracycline regulated wild-type or H115N p53 were generated. H115N was both more potent than wild-type p53 in inducing p53 target genes such as p21 and PIG3 and was also more effective in arresting cells in G1. Unexpectedly, in contrast to wild-type p53, H115N p53 was markedly impaired in causing apoptosis when cells were subjected to DNA damage. Our results indicate that the exonuclease activity and transcriptional activation functions of p53 can be separated. They also extend previous findings showing that cell cycle arrest and apoptosis are separable functions of p53. Finally, these experiments confirm that DNA binding and xonuclease activities are distinct features of the p53 core domain.Fil: Ahn, Jinwoo. University Of Pittsburgh; Estados UnidosFil: Poyurowsky, Masha V.. Columbia University; Estados UnidosFil: Baptiste, Nicole. Columbia University; Estados UnidosFil: Beckerman, Rachel. Columbia University; Estados UnidosFil: Cain, Christine. Columbia University; Estados UnidosFil: Mattia, Melissa. Mount Sinai School of Medicine; Estados UnidosFil: McKinney, Kristine. Harvard Medical School; Estados UnidosFil: Zhou, Jianmin. Columbia University; Estados UnidosFil: Zupnick, Andrew. Columbia University; Estados UnidosFil: Gottifredi, Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Prives, Carol. Columbia University; Estados UnidosLandes Bioscience2009-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/20640Ahn, Jinwoo; Poyurowsky, Masha V.; Baptiste, Nicole; Beckerman, Rachel; Cain, Christine; et al.; Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein; Landes Bioscience; Cell Cycle; 8; 10; 5-2009; 1603-16151538-41011551-4005CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.tandfonline.com/doi/abs/10.4161/cc.8.10.8548info:eu-repo/semantics/altIdentifier/doi/10.4161/cc.8.10.8548info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:36:03Zoai:ri.conicet.gov.ar:11336/20640instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:36:04.248CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
title Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
spellingShingle Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
Ahn, Jinwoo
P53
Apoptosis
Exonuclease Activity
Dna Binding
title_short Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
title_full Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
title_fullStr Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
title_full_unstemmed Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
title_sort Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein
dc.creator.none.fl_str_mv Ahn, Jinwoo
Poyurowsky, Masha V.
Baptiste, Nicole
Beckerman, Rachel
Cain, Christine
Mattia, Melissa
McKinney, Kristine
Zhou, Jianmin
Zupnick, Andrew
Gottifredi, Vanesa
Prives, Carol
author Ahn, Jinwoo
author_facet Ahn, Jinwoo
Poyurowsky, Masha V.
Baptiste, Nicole
Beckerman, Rachel
Cain, Christine
Mattia, Melissa
McKinney, Kristine
Zhou, Jianmin
Zupnick, Andrew
Gottifredi, Vanesa
Prives, Carol
author_role author
author2 Poyurowsky, Masha V.
Baptiste, Nicole
Beckerman, Rachel
Cain, Christine
Mattia, Melissa
McKinney, Kristine
Zhou, Jianmin
Zupnick, Andrew
Gottifredi, Vanesa
Prives, Carol
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv P53
Apoptosis
Exonuclease Activity
Dna Binding
topic P53
Apoptosis
Exonuclease Activity
Dna Binding
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Both sequence-specific DNA binding and exonuclease activities have been mapped to the central conserved core domain of p53. To gain more information about these two activities a series of mutants were generated that changed core domain histidine residues. Of these mutants, only one, H115N p53, showed markedly reduced exonuclease activity (ca. 15% of wild-type). Surprisingly, purified H115N p53 protein was found to be significantly more potent than wild-type p53 in binding to DNA by several criteria including gel mobility shift assay, filter binding and DNase I footprinting. Interestingly as well, non-specific DNA binding by the core domain of H115N p53 is superior to that of wild-type p53. To study H115N p53 in vivo, clones of H1299 cells expressing tetracycline regulated wild-type or H115N p53 were generated. H115N was both more potent than wild-type p53 in inducing p53 target genes such as p21 and PIG3 and was also more effective in arresting cells in G1. Unexpectedly, in contrast to wild-type p53, H115N p53 was markedly impaired in causing apoptosis when cells were subjected to DNA damage. Our results indicate that the exonuclease activity and transcriptional activation functions of p53 can be separated. They also extend previous findings showing that cell cycle arrest and apoptosis are separable functions of p53. Finally, these experiments confirm that DNA binding and xonuclease activities are distinct features of the p53 core domain.
Fil: Ahn, Jinwoo. University Of Pittsburgh; Estados Unidos
Fil: Poyurowsky, Masha V.. Columbia University; Estados Unidos
Fil: Baptiste, Nicole. Columbia University; Estados Unidos
Fil: Beckerman, Rachel. Columbia University; Estados Unidos
Fil: Cain, Christine. Columbia University; Estados Unidos
Fil: Mattia, Melissa. Mount Sinai School of Medicine; Estados Unidos
Fil: McKinney, Kristine. Harvard Medical School; Estados Unidos
Fil: Zhou, Jianmin. Columbia University; Estados Unidos
Fil: Zupnick, Andrew. Columbia University; Estados Unidos
Fil: Gottifredi, Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; Argentina
Fil: Prives, Carol. Columbia University; Estados Unidos
description Both sequence-specific DNA binding and exonuclease activities have been mapped to the central conserved core domain of p53. To gain more information about these two activities a series of mutants were generated that changed core domain histidine residues. Of these mutants, only one, H115N p53, showed markedly reduced exonuclease activity (ca. 15% of wild-type). Surprisingly, purified H115N p53 protein was found to be significantly more potent than wild-type p53 in binding to DNA by several criteria including gel mobility shift assay, filter binding and DNase I footprinting. Interestingly as well, non-specific DNA binding by the core domain of H115N p53 is superior to that of wild-type p53. To study H115N p53 in vivo, clones of H1299 cells expressing tetracycline regulated wild-type or H115N p53 were generated. H115N was both more potent than wild-type p53 in inducing p53 target genes such as p21 and PIG3 and was also more effective in arresting cells in G1. Unexpectedly, in contrast to wild-type p53, H115N p53 was markedly impaired in causing apoptosis when cells were subjected to DNA damage. Our results indicate that the exonuclease activity and transcriptional activation functions of p53 can be separated. They also extend previous findings showing that cell cycle arrest and apoptosis are separable functions of p53. Finally, these experiments confirm that DNA binding and xonuclease activities are distinct features of the p53 core domain.
publishDate 2009
dc.date.none.fl_str_mv 2009-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/20640
Ahn, Jinwoo; Poyurowsky, Masha V.; Baptiste, Nicole; Beckerman, Rachel; Cain, Christine; et al.; Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein; Landes Bioscience; Cell Cycle; 8; 10; 5-2009; 1603-1615
1538-4101
1551-4005
CONICET Digital
CONICET
url http://hdl.handle.net/11336/20640
identifier_str_mv Ahn, Jinwoo; Poyurowsky, Masha V.; Baptiste, Nicole; Beckerman, Rachel; Cain, Christine; et al.; Dissection of the sequence-specific DNA binding and exonuclease activities reveals a superactive yet apoptotically impaired mutant p53 protein; Landes Bioscience; Cell Cycle; 8; 10; 5-2009; 1603-1615
1538-4101
1551-4005
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.tandfonline.com/doi/abs/10.4161/cc.8.10.8548
info:eu-repo/semantics/altIdentifier/doi/10.4161/cc.8.10.8548
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Landes Bioscience
publisher.none.fl_str_mv Landes Bioscience
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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