ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, stru...

Autores
Asensio, Cristian Jorge Alejandro; Sarfield, Simon; Wait, Rob; Crawford, Jane
Año de publicación
2024
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
ZFP36 and ZFP36L1 (L1) are RNA-binding nucleo-cytoplasmic phospho-proteins with zinc-fingers, modulating the decay of AU-rich mRNAs such as those of some inflammatory cytokines. Their individual roles or redundancy in macrophages are unknown. Both are post-translationally modified with multi-site phosphorylation by many kinases and by other PTMs. The impact of all PTMs on their roles, location, mRNA binding, folding, modification code, half-life and interactions are understudied. We aimed to study them in THP1 and HeLa cell lines by subcellular fractionation, immunoprecipitation, immunoblotting, far-WB, dye binding, transfection, radiolabeling and 1D/2D gels, also using enzyme inhibitors and TLR ligands for cell treatments. We studied rZFP36 mutants by kinase assays and ZFP36 by informatic, interactome and MS analysis. As novel PTM, we considered the isomerization in proline-directed phosphosites. By densitometry of their isoforms in gels, results suggested with statistical significance that the hyperphosphorylated forms of L1 and ZFP36 were cytoplasmic but insoluble, interacting with the cytoskeleton. Thus, both distribute in at least 6 locations: cytosol, cytoskeleton, mRNAs, stress-granules, P-bodies and nucleus. Besides,both cellular and rZFP36 isomerize. ZFP36 becomes a model for multi-site phosphorylation and isomerization. L1 levels were different in monocytic and macrophage states, suggesting a role in adifferentiation switch but not in inflammation. ZFP36 was affected by inflammatory signaling but not by macrophage adherence or multinucleation or ribotoxic stress. We visualize a complex rheostatic regulation in which ZFP36 is controlled by interactions with ions, proteins, mRNAs and proteasomes, behaving as a polyanion with electrostatic interactions and disordered regions that can isomerize. More studies are needed to understand their molecular heterogeneity and if they will become drug targets, to fine-tune their many activities without side effects.
Fil: Asensio, Cristian Jorge Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Sarfield, Simon. Imperial College. London Institute Of Medical Sciences.;
Fil: Wait, Rob. Imperial College Of Science And Technology; Reino Unido
Fil: Crawford, Jane. Imperial College Of Science And Technology; Reino Unido
LXIX Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXVI Sociedad Argentina de Fisiología y Asociación Latinoamericana de Ciencias Fisiológicas
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Fisiología
Asociación Latinoamericana de Ciencias Fisiológicas
Materia
RNA-BINDING
INFLAMMATION
PTM
MACROPHAGES
ZFP
KINASES
ISOMERIZATION
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:11336/257727
Acceder
id CONICETDig_c97d5fda8062404d9044f87152dff599
oai_identifier_str oai:ri.conicet.gov.ar:11336/257727
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflifeAsensio, Cristian Jorge AlejandroSarfield, SimonWait, RobCrawford, JaneRNA-BINDINGINFLAMMATIONPTMMACROPHAGESZFPKINASESISOMERIZATIONhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1ZFP36 and ZFP36L1 (L1) are RNA-binding nucleo-cytoplasmic phospho-proteins with zinc-fingers, modulating the decay of AU-rich mRNAs such as those of some inflammatory cytokines. Their individual roles or redundancy in macrophages are unknown. Both are post-translationally modified with multi-site phosphorylation by many kinases and by other PTMs. The impact of all PTMs on their roles, location, mRNA binding, folding, modification code, half-life and interactions are understudied. We aimed to study them in THP1 and HeLa cell lines by subcellular fractionation, immunoprecipitation, immunoblotting, far-WB, dye binding, transfection, radiolabeling and 1D/2D gels, also using enzyme inhibitors and TLR ligands for cell treatments. We studied rZFP36 mutants by kinase assays and ZFP36 by informatic, interactome and MS analysis. As novel PTM, we considered the isomerization in proline-directed phosphosites. By densitometry of their isoforms in gels, results suggested with statistical significance that the hyperphosphorylated forms of L1 and ZFP36 were cytoplasmic but insoluble, interacting with the cytoskeleton. Thus, both distribute in at least 6 locations: cytosol, cytoskeleton, mRNAs, stress-granules, P-bodies and nucleus. Besides,both cellular and rZFP36 isomerize. ZFP36 becomes a model for multi-site phosphorylation and isomerization. L1 levels were different in monocytic and macrophage states, suggesting a role in adifferentiation switch but not in inflammation. ZFP36 was affected by inflammatory signaling but not by macrophage adherence or multinucleation or ribotoxic stress. We visualize a complex rheostatic regulation in which ZFP36 is controlled by interactions with ions, proteins, mRNAs and proteasomes, behaving as a polyanion with electrostatic interactions and disordered regions that can isomerize. More studies are needed to understand their molecular heterogeneity and if they will become drug targets, to fine-tune their many activities without side effects.Fil: Asensio, Cristian Jorge Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Sarfield, Simon. Imperial College. London Institute Of Medical Sciences.;Fil: Wait, Rob. Imperial College Of Science And Technology; Reino UnidoFil: Crawford, Jane. Imperial College Of Science And Technology; Reino UnidoLXIX Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXVI Sociedad Argentina de Fisiología y Asociación Latinoamericana de Ciencias FisiológicasCiudad Autónoma de Buenos AiresArgentinaSociedad Argentina de Investigación ClínicaSociedad Argentina de FisiologíaAsociación Latinoamericana de Ciencias FisiológicasFundación Revista Medicina2024info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/257727ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife; LXIX Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXVI Sociedad Argentina de Fisiología y Asociación Latinoamericana de Ciencias Fisiológicas; Ciudad Autónoma de Buenos Aires; Argentina; 2024; 116-1171669-9106CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.reunionbiociencias.com.ar/wp-content/uploads/2024/12/Revista-Medicina-2024-FINAL.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:25:13Zoai:ri.conicet.gov.ar:11336/257727instacron: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:34982025-10-22 11:25:13.652CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
title ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
spellingShingle ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
Asensio, Cristian Jorge Alejandro
RNA-BINDING
INFLAMMATION
PTM
MACROPHAGES
ZFP
KINASES
ISOMERIZATION
title_short ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
title_full ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
title_fullStr ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
title_full_unstemmed ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
title_sort ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife
dc.creator.none.fl_str_mv Asensio, Cristian Jorge Alejandro
Sarfield, Simon
Wait, Rob
Crawford, Jane
author Asensio, Cristian Jorge Alejandro
author_facet Asensio, Cristian Jorge Alejandro
Sarfield, Simon
Wait, Rob
Crawford, Jane
author_role author
author2 Sarfield, Simon
Wait, Rob
Crawford, Jane
author2_role author
author
author
dc.subject.none.fl_str_mv RNA-BINDING
INFLAMMATION
PTM
MACROPHAGES
ZFP
KINASES
ISOMERIZATION
topic RNA-BINDING
INFLAMMATION
PTM
MACROPHAGES
ZFP
KINASES
ISOMERIZATION
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv ZFP36 and ZFP36L1 (L1) are RNA-binding nucleo-cytoplasmic phospho-proteins with zinc-fingers, modulating the decay of AU-rich mRNAs such as those of some inflammatory cytokines. Their individual roles or redundancy in macrophages are unknown. Both are post-translationally modified with multi-site phosphorylation by many kinases and by other PTMs. The impact of all PTMs on their roles, location, mRNA binding, folding, modification code, half-life and interactions are understudied. We aimed to study them in THP1 and HeLa cell lines by subcellular fractionation, immunoprecipitation, immunoblotting, far-WB, dye binding, transfection, radiolabeling and 1D/2D gels, also using enzyme inhibitors and TLR ligands for cell treatments. We studied rZFP36 mutants by kinase assays and ZFP36 by informatic, interactome and MS analysis. As novel PTM, we considered the isomerization in proline-directed phosphosites. By densitometry of their isoforms in gels, results suggested with statistical significance that the hyperphosphorylated forms of L1 and ZFP36 were cytoplasmic but insoluble, interacting with the cytoskeleton. Thus, both distribute in at least 6 locations: cytosol, cytoskeleton, mRNAs, stress-granules, P-bodies and nucleus. Besides,both cellular and rZFP36 isomerize. ZFP36 becomes a model for multi-site phosphorylation and isomerization. L1 levels were different in monocytic and macrophage states, suggesting a role in adifferentiation switch but not in inflammation. ZFP36 was affected by inflammatory signaling but not by macrophage adherence or multinucleation or ribotoxic stress. We visualize a complex rheostatic regulation in which ZFP36 is controlled by interactions with ions, proteins, mRNAs and proteasomes, behaving as a polyanion with electrostatic interactions and disordered regions that can isomerize. More studies are needed to understand their molecular heterogeneity and if they will become drug targets, to fine-tune their many activities without side effects.
Fil: Asensio, Cristian Jorge Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Sarfield, Simon. Imperial College. London Institute Of Medical Sciences.;
Fil: Wait, Rob. Imperial College Of Science And Technology; Reino Unido
Fil: Crawford, Jane. Imperial College Of Science And Technology; Reino Unido
LXIX Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXVI Sociedad Argentina de Fisiología y Asociación Latinoamericana de Ciencias Fisiológicas
Ciudad Autónoma de Buenos Aires
Argentina
Sociedad Argentina de Investigación Clínica
Sociedad Argentina de Fisiología
Asociación Latinoamericana de Ciencias Fisiológicas
description ZFP36 and ZFP36L1 (L1) are RNA-binding nucleo-cytoplasmic phospho-proteins with zinc-fingers, modulating the decay of AU-rich mRNAs such as those of some inflammatory cytokines. Their individual roles or redundancy in macrophages are unknown. Both are post-translationally modified with multi-site phosphorylation by many kinases and by other PTMs. The impact of all PTMs on their roles, location, mRNA binding, folding, modification code, half-life and interactions are understudied. We aimed to study them in THP1 and HeLa cell lines by subcellular fractionation, immunoprecipitation, immunoblotting, far-WB, dye binding, transfection, radiolabeling and 1D/2D gels, also using enzyme inhibitors and TLR ligands for cell treatments. We studied rZFP36 mutants by kinase assays and ZFP36 by informatic, interactome and MS analysis. As novel PTM, we considered the isomerization in proline-directed phosphosites. By densitometry of their isoforms in gels, results suggested with statistical significance that the hyperphosphorylated forms of L1 and ZFP36 were cytoplasmic but insoluble, interacting with the cytoskeleton. Thus, both distribute in at least 6 locations: cytosol, cytoskeleton, mRNAs, stress-granules, P-bodies and nucleus. Besides,both cellular and rZFP36 isomerize. ZFP36 becomes a model for multi-site phosphorylation and isomerization. L1 levels were different in monocytic and macrophage states, suggesting a role in adifferentiation switch but not in inflammation. ZFP36 was affected by inflammatory signaling but not by macrophage adherence or multinucleation or ribotoxic stress. We visualize a complex rheostatic regulation in which ZFP36 is controlled by interactions with ions, proteins, mRNAs and proteasomes, behaving as a polyanion with electrostatic interactions and disordered regions that can isomerize. More studies are needed to understand their molecular heterogeneity and if they will become drug targets, to fine-tune their many activities without side effects.
publishDate 2024
dc.date.none.fl_str_mv 2024
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/257727
ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife; LXIX Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXVI Sociedad Argentina de Fisiología y Asociación Latinoamericana de Ciencias Fisiológicas; Ciudad Autónoma de Buenos Aires; Argentina; 2024; 116-117
1669-9106
CONICET Digital
CONICET
url http://hdl.handle.net/11336/257727
identifier_str_mv ZFP-36 proteins in monocyte and macrophage inflammatory signaling and differentiation. Model-ling their post-translational modifications, intracellular locations, interactome, structural hetero-geneity and halflife; LXIX Reunión Anual de la Sociedad Argentina de Investigación Clínica; XXVI Sociedad Argentina de Fisiología y Asociación Latinoamericana de Ciencias Fisiológicas; Ciudad Autónoma de Buenos Aires; Argentina; 2024; 116-117
1669-9106
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.reunionbiociencias.com.ar/wp-content/uploads/2024/12/Revista-Medicina-2024-FINAL.pdf
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.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Fundación Revista Medicina
publisher.none.fl_str_mv Fundación Revista Medicina
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
_version_ 1846781798606962688
score 12.982451

Publicaciones similares