Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence
This study continues the search for Plasmodium falciparum transcriptional machinery by isolating DNA binding proteins that could contribute towards discovering new therapeutic targets. The calmodulin gene was thus used as the model, using a 149 bp DNA fragment (PÆ’CAM149) specific for the 5' n...
- Autores:
-
Yara, Erika L
Rojas, María Orfa
- Tipo de recurso:
- Article of journal
- Fecha de publicación:
- 2003
- Institución:
- Universidad Nacional de Colombia
- Repositorio:
- Universidad Nacional de Colombia
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unal.edu.co:unal/22188
- Acceso en línea:
- https://repositorio.unal.edu.co/handle/unal/22188
http://bdigital.unal.edu.co/13222/
- Palabra clave:
- malaria
Plasmodium falciparum
maquinaria
transcripción
factores de transcripción
malaria
Plasmodium falciparum
machinery
transcription
transcription factors
- Rights
- closedAccess
- License
- Atribución-NoComercial 4.0 Internacional
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Universidad Nacional de Colombia |
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|
dc.title.spa.fl_str_mv |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence |
title |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence |
spellingShingle |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence malaria Plasmodium falciparum maquinaria transcripción factores de transcripción malaria Plasmodium falciparum machinery transcription transcription factors |
title_short |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence |
title_full |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence |
title_fullStr |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence |
title_full_unstemmed |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence |
title_sort |
Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence |
dc.creator.fl_str_mv |
Yara, Erika L Rojas, María Orfa |
dc.contributor.author.spa.fl_str_mv |
Yara, Erika L Rojas, María Orfa |
dc.subject.proposal.spa.fl_str_mv |
malaria Plasmodium falciparum maquinaria transcripción factores de transcripción malaria Plasmodium falciparum machinery transcription transcription factors |
topic |
malaria Plasmodium falciparum maquinaria transcripción factores de transcripción malaria Plasmodium falciparum machinery transcription transcription factors |
description |
This study continues the search for Plasmodium falciparum transcriptional machinery by isolating DNA binding proteins that could contribute towards discovering new therapeutic targets. The calmodulin gene was thus used as the model, using a 149 bp DNA fragment (PÆ’CAM149) specific for the 5' non-translated region. Partially purified proteins derived from total parasite nuclear extract were used to scale the PÆ’CAM149-biotina-streptavidin-magnesphere DNA affinity microsystem previously developed for capturing those proteins specifically binding to the DNA PÆ’CAM149 sequence. The scaling allowed the protein to be recycled three times through chromatographic matrix and determine which proteins remained bound to the system, indicating specific interaction with the calmodulin gene (PÆ’CAM149) promoter sequence. It also enabled obtaining the quantity of protein necessary for sequencing. Other assays allowed binding activity to be tracked by using eukaryotic transcription factor consensus sequences (CREB, OCT1, MIG, GATA), leading to selecting a 54 kDa protein whose possible identity corresponds to the E4BP4 transcription factor or related sequences. Problems regarding quantity were overcome in this work. Binding specificity was ascertained and it was determined that the 54 kDa protein had an intrinsic modification impeding its amino terminal being sequenced. These assays showed that CREB, or related transcription factors, form part of the parasite's transcriptional machinery. Key words: malaria; Plasmodium falciparum; machinery; transcription; transcription factors |
publishDate |
2003 |
dc.date.issued.spa.fl_str_mv |
2003 |
dc.date.accessioned.spa.fl_str_mv |
2019-06-25T20:33:52Z |
dc.date.available.spa.fl_str_mv |
2019-06-25T20:33:52Z |
dc.type.spa.fl_str_mv |
Artículo de revista |
dc.type.coar.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_6501 |
dc.type.coarversion.spa.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
dc.type.content.spa.fl_str_mv |
Text |
dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/ART |
format |
http://purl.org/coar/resource_type/c_6501 |
status_str |
publishedVersion |
dc.identifier.uri.none.fl_str_mv |
https://repositorio.unal.edu.co/handle/unal/22188 |
dc.identifier.eprints.spa.fl_str_mv |
http://bdigital.unal.edu.co/13222/ |
url |
https://repositorio.unal.edu.co/handle/unal/22188 http://bdigital.unal.edu.co/13222/ |
dc.language.iso.spa.fl_str_mv |
spa |
language |
spa |
dc.relation.spa.fl_str_mv |
http://revistas.unal.edu.co/index.php/biotecnologia/article/view/595 |
dc.relation.ispartof.spa.fl_str_mv |
Universidad Nacional de Colombia Revistas electrónicas UN Revista Colombiana de Biotecnología Revista Colombiana de Biotecnología |
dc.relation.ispartofseries.none.fl_str_mv |
Revista Colombiana de Biotecnología; Vol. 5, núm. 1 (2003); 82-95 1909-8758 0123-3475 |
dc.relation.references.spa.fl_str_mv |
Yara, Erika L and Rojas, María Orfa (2003) Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence. Revista Colombiana de Biotecnología; Vol. 5, núm. 1 (2003); 82-95 1909-8758 0123-3475 . |
dc.rights.spa.fl_str_mv |
Derechos reservados - Universidad Nacional de Colombia |
dc.rights.coar.fl_str_mv |
http://purl.org/coar/access_right/c_14cb |
dc.rights.license.spa.fl_str_mv |
Atribución-NoComercial 4.0 Internacional |
dc.rights.uri.spa.fl_str_mv |
http://creativecommons.org/licenses/by-nc/4.0/ |
dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/closedAccess |
rights_invalid_str_mv |
Atribución-NoComercial 4.0 Internacional Derechos reservados - Universidad Nacional de Colombia http://creativecommons.org/licenses/by-nc/4.0/ http://purl.org/coar/access_right/c_14cb |
eu_rights_str_mv |
closedAccess |
dc.publisher.spa.fl_str_mv |
Universidad Nacional de Colombia |
institution |
Universidad Nacional de Colombia |
repository.name.fl_str_mv |
Repositorio Institucional Universidad Nacional de Colombia |
repository.mail.fl_str_mv |
repositorio_nal@unal.edu.co |
_version_ |
1814089231144845312 |
spelling |
Atribución-NoComercial 4.0 InternacionalDerechos reservados - Universidad Nacional de Colombiahttp://creativecommons.org/licenses/by-nc/4.0/info:eu-repo/semantics/closedAccesshttp://purl.org/coar/access_right/c_14cbYara, Erika Lf65ac70f-f39f-43c2-a643-5e5a35fc8b25300Rojas, María Orfa3dd3433e-ec23-4cbb-bd0c-628800e0aff33002019-06-25T20:33:52Z2019-06-25T20:33:52Z2003https://repositorio.unal.edu.co/handle/unal/22188http://bdigital.unal.edu.co/13222/This study continues the search for Plasmodium falciparum transcriptional machinery by isolating DNA binding proteins that could contribute towards discovering new therapeutic targets. The calmodulin gene was thus used as the model, using a 149 bp DNA fragment (PÆ’CAM149) specific for the 5' non-translated region. Partially purified proteins derived from total parasite nuclear extract were used to scale the PÆ’CAM149-biotina-streptavidin-magnesphere DNA affinity microsystem previously developed for capturing those proteins specifically binding to the DNA PÆ’CAM149 sequence. The scaling allowed the protein to be recycled three times through chromatographic matrix and determine which proteins remained bound to the system, indicating specific interaction with the calmodulin gene (PÆ’CAM149) promoter sequence. It also enabled obtaining the quantity of protein necessary for sequencing. Other assays allowed binding activity to be tracked by using eukaryotic transcription factor consensus sequences (CREB, OCT1, MIG, GATA), leading to selecting a 54 kDa protein whose possible identity corresponds to the E4BP4 transcription factor or related sequences. Problems regarding quantity were overcome in this work. Binding specificity was ascertained and it was determined that the 54 kDa protein had an intrinsic modification impeding its amino terminal being sequenced. These assays showed that CREB, or related transcription factors, form part of the parasite's transcriptional machinery. Key words: malaria; Plasmodium falciparum; machinery; transcription; transcription factorsEn este trabajo se sigue la búsqueda de la maquinaria transcripcional de Plasmodium falciparum, aislando proteínas de unión a DNA, que podrían contribuir en un futuro a encontrar nuevos blancos terapéuticos. Con este fin se tomó como modelo el gen de calmodulina, utilizando específicamente un fragmento de DNA de 149 pb (PfCAM149) de la región 5' no traducida. Proteínas parcialmente purificadas provenientes del extracto nuclear total del parásito, se usaron para escalar el microsistema de afinidad DNA PfCAM149-biotina- estreptavidina- magnesferas, previamente desarrollado, para capturar aquellas proteínas de unión específica a la secuencia de DNA PfCAM149. El escalamiento permitió reci-clar tres veces la proteína a través de la matriz cromatográfica y determinar cuáles proteínas permanecían unidas al sistema, indicando interacción específica a la secuencia del promotor del gen de calmodulina (PfCAM149); además, condujo a obtener la cantidad de proteína necesaria para secuenciación. Por otra parte, se realizaron ensayos para rastrear la actividad de unión con secuencias consenso para factores de transcripción eucariotes (CREB, OCT1, MIG y GATA), que permitieron elegir una proteína de 54 kDa, cuya posible identidad corresponde al factor de transcripción E4BP4 o secuencias relacionadas. Con este trabajo se superaron problemas de cantidad, se comprobó la especificidad de la unión y se determinó que la proteína de 54 kDa tenía una modificación intrínseca que impidió la secuenciación de su extremo aminoterminal. Los ensayos realizados evidencian que el factor de trascripción CREB, o factores de transcripción relacionados forman parte de la maquinaria transcripcional del parásito. Palabras claves: malaria; Plasmodium falciparum; maquinaria; transcripción; factores de transcripción.spaUniversidad Nacional de Colombiahttp://revistas.unal.edu.co/index.php/biotecnologia/article/view/595Universidad Nacional de Colombia Revistas electrónicas UN Revista Colombiana de BiotecnologíaRevista Colombiana de BiotecnologíaRevista Colombiana de Biotecnología; Vol. 5, núm. 1 (2003); 82-95 1909-8758 0123-3475Yara, Erika L and Rojas, María Orfa (2003) Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequence. Revista Colombiana de Biotecnología; Vol. 5, núm. 1 (2003); 82-95 1909-8758 0123-3475 .Isolating and identifying proteins binding to a specific plasmodium falciparum calmodulin gene promotor sequenceArtículo de revistainfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501http://purl.org/coar/resource_type/c_2df8fbb1http://purl.org/coar/version/c_970fb48d4fbd8a85Texthttp://purl.org/redcol/resource_type/ARTmalariaPlasmodium falciparummaquinariatranscripciónfactores de transcripciónmalariaPlasmodium falciparummachinerytranscriptiontranscription factorsunal/22188oai:repositorio.unal.edu.co:unal/221882021-04-23 10:13:27.353Repositorio Institucional Universidad Nacional de Colombiarepositorio_nal@unal.edu.co |