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
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/191956
Acceder
id CONICETDig_d9e262749800a10a437824e500464005
oai_identifier_str oai:ri.conicet.gov.ar:11336/191956
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
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 Técnicas2025-09-29T10:41:39Zoai: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:34982025-09-29 10:41:39.892CONICET 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
_version_ 1844614447878897664
score 13.070432

Publicaciones similares