Mapping of the Self-Interaction Domains in the Simian Immunodeficiency Virus Gag Polyprotein

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 GagΔp6) assembled in vitro into spherical particles. Based on the full multimerization capacity of...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Rauddi, María L., Mac Donald, Cecilia L., Affranchino, José L., González, Silvia A.
Formato:	Artículo
Lenguaje:	Inglés
Publicado:	Universidad de Belgrano - Facultad de Ciencias Exactas y Naturales - Proyectos de Investigación 2014
Materias:	Simian Immunodeficiency Virus Gag Polyprotein Self-Interaction Domains Dominios libre interacción Gag poliproteína Virus de Inmunodeficiencia de simios
Acceso en línea:	http://repositorio.ub.edu.ar/handle/123456789/2713
Aporte de:	Repositorio Institucional - Universidad de Belgrano (UB) de Universidad de Belgrano

id	I36-R142-123456789-2713
record_format	dspace
institution	Universidad de Belgrano
institution_str	I-36
repository_str	R-142
collection	Repositorio Institucional - Universidad de Belgrano (UB)
language	Inglés
topic	Simian Immunodeficiency Virus Gag Polyprotein Self-Interaction Domains Dominios libre interacción Gag poliproteína Virus de Inmunodeficiencia de simios
spellingShingle	Simian Immunodeficiency Virus Gag Polyprotein Self-Interaction Domains Dominios libre interacción Gag poliproteína Virus de Inmunodeficiencia de simios Rauddi, María L. Mac Donald, Cecilia L. Affranchino, José L. González, Silvia A. Mapping of the Self-Interaction Domains in the Simian Immunodeficiency Virus Gag Polyprotein
topic_facet	Simian Immunodeficiency Virus Gag Polyprotein Self-Interaction Domains Dominios libre interacción Gag poliproteína Virus de Inmunodeficiencia de simios
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 GagΔp6) assembled in vitro into spherical particles. Based on the full multimerization capacity of SIV GagΔp6, 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 GST-SIVGagΔp6. Removal of the nucleocapsid (NC) domain from Gag impaired its ability to interact with GST-SIVGagΔp6. 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 vitrowith GST-SIVGagΔp6. Notably, a small Gag subdomain containing the C-terminal third of the CA and the entire NC not only bound to GST-SIVGagΔp6 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-SIVGagΔp6 with efficiencies representing 20% and 40%, respectively, of the wild-type value, whereas the CA failed to bind to GST-SIVGagΔp6, despite being capable of self-associating into multimeric complexes.
format	Article
author	Rauddi, María L. Mac Donald, Cecilia L. Affranchino, José L. González, Silvia A.
author_facet	Rauddi, María L. Mac Donald, Cecilia L. Affranchino, José L. González, Silvia A.
author_sort	Rauddi, María L.
title	Mapping of the Self-Interaction Domains in the Simian Immunodeficiency Virus Gag Polyprotein
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
publisher	Universidad de Belgrano - Facultad de Ciencias Exactas y Naturales - Proyectos de Investigación
publishDate	2014
url	http://repositorio.ub.edu.ar/handle/123456789/2713
work_keys_str_mv	AT rauddimarial mappingoftheselfinteractiondomainsinthesimianimmunodeficiencyvirusgagpolyprotein AT macdonaldcecilial mappingoftheselfinteractiondomainsinthesimianimmunodeficiencyvirusgagpolyprotein AT affranchinojosel mappingoftheselfinteractiondomainsinthesimianimmunodeficiencyvirusgagpolyprotein AT gonzalezsilviaa mappingoftheselfinteractiondomainsinthesimianimmunodeficiencyvirusgagpolyprotein
bdutipo_str	Repositorios
_version_	1764820529225662467

Mapping of the Self-Interaction Domains in the Simian Immunodeficiency Virus Gag Polyprotein

Ejemplares similares