Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein
- Autores
- Rauddi, Maria Luisa; Mac Donald, Cecilia L.; Affranchino, Jose Luis; Gonzalez, Silvia Adriana
- Año de publicación
- 2011
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- To gain a better understanding of the assembly process in simian immunodeficiency virus (SIV), we first established the conditions under which recombinant SIV Gag lacking the C-terminal p6 domain (SIV GagDp6) assembled in vitro into spherical particles. Based on the full multimerization capacity of SIV GagDp6, and to identify the Gag sequences involved in homotypic interactions, we next developed a pull-down assay in which a panel of histidine-tagged SIV Gag truncation mutants was tested for its ability to associate in vitro with GSTSIVGagDp6. Removal of the nucleocapsid (NC) domain from Gag impaired its ability to interact with GSTSIVGagDp6. However, this Gag mutant consisting of the matrix (MA) and capsid (CA) domains still retained 50% of the wild-type binding activity. Truncation of SIV Gag from its N-terminus yielded markedly different results. The Gag region consisting of the CA and NC was significantly more efficient than wild-type Gag at interacting in vitro with GST-SIVGagDp6. Notably, a small Gag subdomain containing the C-terminal third of the CA and the entire NC not only bound to GST-SIVGagDp6 in vitro at wild-type levels, but also associated in vivo with full-length Gag and was recruited into extracellular particles. Interestingly, when the mature Gag products were analyzed, the MA and NC interacted with GST-SIVGagDp6 with efficiencies representing 20% and 40%, respectively, of the wild-type value, whereas the CA failed to bind to GST-SIVGagDp6, despite being capable of self-associating into multimeric complexes.
Fil: Rauddi, Maria Luisa. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Mac Donald, Cecilia L.. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina
Fil: Affranchino, Jose Luis. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gonzalez, Silvia Adriana. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina - Materia
-
SIV
GAG POLYPROTEIN
VIRUS ASSEMBLY
LENTIVIRUSES - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/191956
Ver los metadatos del registro completo
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network_name_str |
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spelling |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyproteinRauddi, Maria LuisaMac Donald, Cecilia L.Affranchino, Jose LuisGonzalez, Silvia AdrianaSIVGAG POLYPROTEINVIRUS ASSEMBLYLENTIVIRUSEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1https://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1To gain a better understanding of the assembly process in simian immunodeficiency virus (SIV), we first established the conditions under which recombinant SIV Gag lacking the C-terminal p6 domain (SIV GagDp6) assembled in vitro into spherical particles. Based on the full multimerization capacity of SIV GagDp6, and to identify the Gag sequences involved in homotypic interactions, we next developed a pull-down assay in which a panel of histidine-tagged SIV Gag truncation mutants was tested for its ability to associate in vitro with GSTSIVGagDp6. Removal of the nucleocapsid (NC) domain from Gag impaired its ability to interact with GSTSIVGagDp6. However, this Gag mutant consisting of the matrix (MA) and capsid (CA) domains still retained 50% of the wild-type binding activity. Truncation of SIV Gag from its N-terminus yielded markedly different results. The Gag region consisting of the CA and NC was significantly more efficient than wild-type Gag at interacting in vitro with GST-SIVGagDp6. Notably, a small Gag subdomain containing the C-terminal third of the CA and the entire NC not only bound to GST-SIVGagDp6 in vitro at wild-type levels, but also associated in vivo with full-length Gag and was recruited into extracellular particles. Interestingly, when the mature Gag products were analyzed, the MA and NC interacted with GST-SIVGagDp6 with efficiencies representing 20% and 40%, respectively, of the wild-type value, whereas the CA failed to bind to GST-SIVGagDp6, despite being capable of self-associating into multimeric complexes.Fil: Rauddi, Maria Luisa. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mac Donald, Cecilia L.. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; ArgentinaFil: Affranchino, Jose Luis. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gonzalez, Silvia Adriana. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaMary Ann Liebert2011-03-10info: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/11336/191956Rauddi, Maria Luisa; Mac Donald, Cecilia L. ; Affranchino, Jose Luis; Gonzalez, Silvia Adriana; Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein; Mary Ann Liebert; Aids Research and Human Retroviruses; 27; 3; 10-3-2011; 303-3160889-22291931-8405CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.liebertpub.com/doi/10.1089/aid.2010.0137info:eu-repo/semantics/altIdentifier/doi/10.1089/aid.2010.0137info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:10:02Zoai:ri.conicet.gov.ar:11336/191956instacron: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:34982025-09-03 10:10:03.101CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein |
title |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein |
spellingShingle |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein Rauddi, Maria Luisa SIV GAG POLYPROTEIN VIRUS ASSEMBLY LENTIVIRUSES |
title_short |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein |
title_full |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein |
title_fullStr |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein |
title_full_unstemmed |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein |
title_sort |
Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein |
dc.creator.none.fl_str_mv |
Rauddi, Maria Luisa Mac Donald, Cecilia L. Affranchino, Jose Luis Gonzalez, Silvia Adriana |
author |
Rauddi, Maria Luisa |
author_facet |
Rauddi, Maria Luisa Mac Donald, Cecilia L. Affranchino, Jose Luis Gonzalez, Silvia Adriana |
author_role |
author |
author2 |
Mac Donald, Cecilia L. Affranchino, Jose Luis Gonzalez, Silvia Adriana |
author2_role |
author author author |
dc.subject.none.fl_str_mv |
SIV GAG POLYPROTEIN VIRUS ASSEMBLY LENTIVIRUSES |
topic |
SIV GAG POLYPROTEIN VIRUS ASSEMBLY LENTIVIRUSES |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
To gain a better understanding of the assembly process in simian immunodeficiency virus (SIV), we first established the conditions under which recombinant SIV Gag lacking the C-terminal p6 domain (SIV GagDp6) assembled in vitro into spherical particles. Based on the full multimerization capacity of SIV GagDp6, and to identify the Gag sequences involved in homotypic interactions, we next developed a pull-down assay in which a panel of histidine-tagged SIV Gag truncation mutants was tested for its ability to associate in vitro with GSTSIVGagDp6. Removal of the nucleocapsid (NC) domain from Gag impaired its ability to interact with GSTSIVGagDp6. However, this Gag mutant consisting of the matrix (MA) and capsid (CA) domains still retained 50% of the wild-type binding activity. Truncation of SIV Gag from its N-terminus yielded markedly different results. The Gag region consisting of the CA and NC was significantly more efficient than wild-type Gag at interacting in vitro with GST-SIVGagDp6. Notably, a small Gag subdomain containing the C-terminal third of the CA and the entire NC not only bound to GST-SIVGagDp6 in vitro at wild-type levels, but also associated in vivo with full-length Gag and was recruited into extracellular particles. Interestingly, when the mature Gag products were analyzed, the MA and NC interacted with GST-SIVGagDp6 with efficiencies representing 20% and 40%, respectively, of the wild-type value, whereas the CA failed to bind to GST-SIVGagDp6, despite being capable of self-associating into multimeric complexes. Fil: Rauddi, Maria Luisa. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Mac Donald, Cecilia L.. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina Fil: Affranchino, Jose Luis. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Gonzalez, Silvia Adriana. Universidad de Belgrano. Facultad de Ciencias Exactas y Naturales. Laboratorio de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina |
description |
To gain a better understanding of the assembly process in simian immunodeficiency virus (SIV), we first established the conditions under which recombinant SIV Gag lacking the C-terminal p6 domain (SIV GagDp6) assembled in vitro into spherical particles. Based on the full multimerization capacity of SIV GagDp6, and to identify the Gag sequences involved in homotypic interactions, we next developed a pull-down assay in which a panel of histidine-tagged SIV Gag truncation mutants was tested for its ability to associate in vitro with GSTSIVGagDp6. Removal of the nucleocapsid (NC) domain from Gag impaired its ability to interact with GSTSIVGagDp6. However, this Gag mutant consisting of the matrix (MA) and capsid (CA) domains still retained 50% of the wild-type binding activity. Truncation of SIV Gag from its N-terminus yielded markedly different results. The Gag region consisting of the CA and NC was significantly more efficient than wild-type Gag at interacting in vitro with GST-SIVGagDp6. Notably, a small Gag subdomain containing the C-terminal third of the CA and the entire NC not only bound to GST-SIVGagDp6 in vitro at wild-type levels, but also associated in vivo with full-length Gag and was recruited into extracellular particles. Interestingly, when the mature Gag products were analyzed, the MA and NC interacted with GST-SIVGagDp6 with efficiencies representing 20% and 40%, respectively, of the wild-type value, whereas the CA failed to bind to GST-SIVGagDp6, despite being capable of self-associating into multimeric complexes. |
publishDate |
2011 |
dc.date.none.fl_str_mv |
2011-03-10 |
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/11336/191956 Rauddi, Maria Luisa; Mac Donald, Cecilia L. ; Affranchino, Jose Luis; Gonzalez, Silvia Adriana; Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein; Mary Ann Liebert; Aids Research and Human Retroviruses; 27; 3; 10-3-2011; 303-316 0889-2229 1931-8405 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/191956 |
identifier_str_mv |
Rauddi, Maria Luisa; Mac Donald, Cecilia L. ; Affranchino, Jose Luis; Gonzalez, Silvia Adriana; Mapping of the self-interaction domains in the simian immunodeficiency virus gag polyprotein; Mary Ann Liebert; Aids Research and Human Retroviruses; 27; 3; 10-3-2011; 303-316 0889-2229 1931-8405 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.liebertpub.com/doi/10.1089/aid.2010.0137 info:eu-repo/semantics/altIdentifier/doi/10.1089/aid.2010.0137 |
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 application/pdf |
dc.publisher.none.fl_str_mv |
Mary Ann Liebert |
publisher.none.fl_str_mv |
Mary Ann Liebert |
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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1842270104363991040 |
score |
13.13397 |