A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei

Autores
Natalini, Paola Marisel; Mateos, Melina Valeria; Ilincheta, Monica Graciela; Giusto, Norma Maria
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In this work, we describe a selective light-dependent distribution of the lipid kinase 1,2-diacylglycerol kinase (EC 2.7.1.107, DAGK) and the phosphorylated protein kinase C alpha (pPKCalpha) in a nuclear fraction of photoreceptor cells from bovine retinas. A nuclear fraction enriched in small nuclei from photoreceptor cells (PNF), was obtained when a modified nuclear isolation protocol developed by our laboratory was used. We measured and compared DAGK activity as phosphatidic acid (PA) formation in PNF obtained from retinas exposed to light and in retinas kept in darkness using [gamma-32P]ATP or [3H]DAG. In the absence of exogenous substrates and detergents, no changes in DAGK activity were observed. However, when DAGK activity assays were performed in the presence of exogenous substrates, such as stearoyl arachidonoyl glycerol (SAG) or dioleoyl glycerol (DOG), and different detergents (used to make different DAGK isoforms evident), we observed significant light effects on DAGK activity, suggesting the presence of several DAGK isoforms in PNF. Under conditions favoring DAGKzeta activity (DOG, Triton X-100, dioleoyl phosphatidylserine and R59022) we observed an increase in PA formation in PNF from retinas exposed to light with respect to those exposed to darkness. In contrast, under conditions favoring DAGKepsilon (SAG, octylglucoside and R59022) we observed a decrease in its activity. These results suggest different physiological roles of the above-mentioned DAGK isoforms. Western blot analysis showed that whereas light stimulation of bovine retinas increases DAGKzeta nuclear content, it decreases DAGKepsilon and DAGKbeta content in PNF. The role of PIP2-phospholipase C in light-stimulated DAGK activity was demonstrated using U73122. Light was also observed to induce enhanced pPKCalpha content in PNF. The selective distribution of DAGKzeta and epsilon in PNF could be a light-dependent mechanism that in vertebrate retina promotes selective DAG removal and PKC regulation.
Fil: Natalini, Paola Marisel. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
Fil: Mateos, Melina Valeria. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
Fil: Ilincheta, Monica Graciela. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
Fil: Giusto, Norma Maria. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
Materia
Diacylglycerol Kinase
Protein Kinase C
Bovine Photoreceptor Nuclei
Light-Dependent Distribution
Retina
Lipid Second Messengers
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/4504
Acceder
id CONICETDig_8d5247951a71898608dd1f7f1238c3dc
oai_identifier_str oai:ri.conicet.gov.ar:11336/4504
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nucleiNatalini, Paola MariselMateos, Melina ValeriaIlincheta, Monica GracielaGiusto, Norma MariaDiacylglycerol KinaseProtein Kinase CBovine Photoreceptor NucleiLight-Dependent DistributionRetinaLipid Second Messengershttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1In this work, we describe a selective light-dependent distribution of the lipid kinase 1,2-diacylglycerol kinase (EC 2.7.1.107, DAGK) and the phosphorylated protein kinase C alpha (pPKCalpha) in a nuclear fraction of photoreceptor cells from bovine retinas. A nuclear fraction enriched in small nuclei from photoreceptor cells (PNF), was obtained when a modified nuclear isolation protocol developed by our laboratory was used. We measured and compared DAGK activity as phosphatidic acid (PA) formation in PNF obtained from retinas exposed to light and in retinas kept in darkness using [gamma-32P]ATP or [3H]DAG. In the absence of exogenous substrates and detergents, no changes in DAGK activity were observed. However, when DAGK activity assays were performed in the presence of exogenous substrates, such as stearoyl arachidonoyl glycerol (SAG) or dioleoyl glycerol (DOG), and different detergents (used to make different DAGK isoforms evident), we observed significant light effects on DAGK activity, suggesting the presence of several DAGK isoforms in PNF. Under conditions favoring DAGKzeta activity (DOG, Triton X-100, dioleoyl phosphatidylserine and R59022) we observed an increase in PA formation in PNF from retinas exposed to light with respect to those exposed to darkness. In contrast, under conditions favoring DAGKepsilon (SAG, octylglucoside and R59022) we observed a decrease in its activity. These results suggest different physiological roles of the above-mentioned DAGK isoforms. Western blot analysis showed that whereas light stimulation of bovine retinas increases DAGKzeta nuclear content, it decreases DAGKepsilon and DAGKbeta content in PNF. The role of PIP2-phospholipase C in light-stimulated DAGK activity was demonstrated using U73122. Light was also observed to induce enhanced pPKCalpha content in PNF. The selective distribution of DAGKzeta and epsilon in PNF could be a light-dependent mechanism that in vertebrate retina promotes selective DAG removal and PKC regulation.Fil: Natalini, Paola Marisel. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); ArgentinaFil: Mateos, Melina Valeria. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); ArgentinaFil: Ilincheta, Monica Graciela. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); ArgentinaFil: Giusto, Norma Maria. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); ArgentinaAcademic Press Ltd-elsevier Science Ltd2014-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/4504Natalini, Paola Marisel; Mateos, Melina Valeria; Ilincheta, Monica Graciela; Giusto, Norma Maria; A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei; Academic Press Ltd-elsevier Science Ltd; Experimental Eye Research; 125; 6-2014; 142-1550014-4835enginfo:eu-repo/semantics/altIdentifier/doi/info:eu-repo/semantics/altIdentifier/url/http://www.ncbi.nlm.nih.gov/pubmed/24950064info:eu-repo/semantics/altIdentifier/doi/10.1016/j.exer.2014.06.007info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:11:36Zoai:ri.conicet.gov.ar:11336/4504instacron: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:34982025-09-10 13:11:36.863CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
title A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
spellingShingle A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
Natalini, Paola Marisel
Diacylglycerol Kinase
Protein Kinase C
Bovine Photoreceptor Nuclei
Light-Dependent Distribution
Retina
Lipid Second Messengers
title_short A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
title_full A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
title_fullStr A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
title_full_unstemmed A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
title_sort A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei
dc.creator.none.fl_str_mv Natalini, Paola Marisel
Mateos, Melina Valeria
Ilincheta, Monica Graciela
Giusto, Norma Maria
author Natalini, Paola Marisel
author_facet Natalini, Paola Marisel
Mateos, Melina Valeria
Ilincheta, Monica Graciela
Giusto, Norma Maria
author_role author
author2 Mateos, Melina Valeria
Ilincheta, Monica Graciela
Giusto, Norma Maria
author2_role author
author
author
dc.subject.none.fl_str_mv Diacylglycerol Kinase
Protein Kinase C
Bovine Photoreceptor Nuclei
Light-Dependent Distribution
Retina
Lipid Second Messengers
topic Diacylglycerol Kinase
Protein Kinase C
Bovine Photoreceptor Nuclei
Light-Dependent Distribution
Retina
Lipid Second Messengers
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In this work, we describe a selective light-dependent distribution of the lipid kinase 1,2-diacylglycerol kinase (EC 2.7.1.107, DAGK) and the phosphorylated protein kinase C alpha (pPKCalpha) in a nuclear fraction of photoreceptor cells from bovine retinas. A nuclear fraction enriched in small nuclei from photoreceptor cells (PNF), was obtained when a modified nuclear isolation protocol developed by our laboratory was used. We measured and compared DAGK activity as phosphatidic acid (PA) formation in PNF obtained from retinas exposed to light and in retinas kept in darkness using [gamma-32P]ATP or [3H]DAG. In the absence of exogenous substrates and detergents, no changes in DAGK activity were observed. However, when DAGK activity assays were performed in the presence of exogenous substrates, such as stearoyl arachidonoyl glycerol (SAG) or dioleoyl glycerol (DOG), and different detergents (used to make different DAGK isoforms evident), we observed significant light effects on DAGK activity, suggesting the presence of several DAGK isoforms in PNF. Under conditions favoring DAGKzeta activity (DOG, Triton X-100, dioleoyl phosphatidylserine and R59022) we observed an increase in PA formation in PNF from retinas exposed to light with respect to those exposed to darkness. In contrast, under conditions favoring DAGKepsilon (SAG, octylglucoside and R59022) we observed a decrease in its activity. These results suggest different physiological roles of the above-mentioned DAGK isoforms. Western blot analysis showed that whereas light stimulation of bovine retinas increases DAGKzeta nuclear content, it decreases DAGKepsilon and DAGKbeta content in PNF. The role of PIP2-phospholipase C in light-stimulated DAGK activity was demonstrated using U73122. Light was also observed to induce enhanced pPKCalpha content in PNF. The selective distribution of DAGKzeta and epsilon in PNF could be a light-dependent mechanism that in vertebrate retina promotes selective DAG removal and PKC regulation.
Fil: Natalini, Paola Marisel. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
Fil: Mateos, Melina Valeria. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
Fil: Ilincheta, Monica Graciela. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
Fil: Giusto, Norma Maria. Universidad Nacional del Sur; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET Bahía Blanca. Instituto de Investigaciones Bioquímicas Bahía Blanca (i); Argentina
description In this work, we describe a selective light-dependent distribution of the lipid kinase 1,2-diacylglycerol kinase (EC 2.7.1.107, DAGK) and the phosphorylated protein kinase C alpha (pPKCalpha) in a nuclear fraction of photoreceptor cells from bovine retinas. A nuclear fraction enriched in small nuclei from photoreceptor cells (PNF), was obtained when a modified nuclear isolation protocol developed by our laboratory was used. We measured and compared DAGK activity as phosphatidic acid (PA) formation in PNF obtained from retinas exposed to light and in retinas kept in darkness using [gamma-32P]ATP or [3H]DAG. In the absence of exogenous substrates and detergents, no changes in DAGK activity were observed. However, when DAGK activity assays were performed in the presence of exogenous substrates, such as stearoyl arachidonoyl glycerol (SAG) or dioleoyl glycerol (DOG), and different detergents (used to make different DAGK isoforms evident), we observed significant light effects on DAGK activity, suggesting the presence of several DAGK isoforms in PNF. Under conditions favoring DAGKzeta activity (DOG, Triton X-100, dioleoyl phosphatidylserine and R59022) we observed an increase in PA formation in PNF from retinas exposed to light with respect to those exposed to darkness. In contrast, under conditions favoring DAGKepsilon (SAG, octylglucoside and R59022) we observed a decrease in its activity. These results suggest different physiological roles of the above-mentioned DAGK isoforms. Western blot analysis showed that whereas light stimulation of bovine retinas increases DAGKzeta nuclear content, it decreases DAGKepsilon and DAGKbeta content in PNF. The role of PIP2-phospholipase C in light-stimulated DAGK activity was demonstrated using U73122. Light was also observed to induce enhanced pPKCalpha content in PNF. The selective distribution of DAGKzeta and epsilon in PNF could be a light-dependent mechanism that in vertebrate retina promotes selective DAG removal and PKC regulation.
publishDate 2014
dc.date.none.fl_str_mv 2014-06
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/11336/4504
Natalini, Paola Marisel; Mateos, Melina Valeria; Ilincheta, Monica Graciela; Giusto, Norma Maria; A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei; Academic Press Ltd-elsevier Science Ltd; Experimental Eye Research; 125; 6-2014; 142-155
0014-4835
url http://hdl.handle.net/11336/4504
identifier_str_mv Natalini, Paola Marisel; Mateos, Melina Valeria; Ilincheta, Monica Graciela; Giusto, Norma Maria; A novel light-dependent activation of DAGK and PKC in bovine photoreceptor nuclei; Academic Press Ltd-elsevier Science Ltd; Experimental Eye Research; 125; 6-2014; 142-155
0014-4835
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/
info:eu-repo/semantics/altIdentifier/url/http://www.ncbi.nlm.nih.gov/pubmed/24950064
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.exer.2014.06.007
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Academic Press Ltd-elsevier Science Ltd
publisher.none.fl_str_mv Academic Press Ltd-elsevier Science Ltd
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 12.993085

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