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Title: Ubiquitin-fusion as a strategy to modulate protein half-life: A3G antiviral activity revisited

Abstract

The human APOBEC3G (A3G) is a potent inhibitor of HIV-1 replication and its activity is suppressed by HIV-1 virion infectivity factor (Vif). Vif neutralizes A3G mainly by inducing its degradation in the proteasome and blocking its incorporation into HIV-1 virions. Assessing the time needed for A3G incorporation into virions is, therefore, important to determine how quickly Vif must act to induce its degradation. We show that modelling the intracellular half-life of A3G can induce its Vif-independent targeting to the ubiquitin-proteasome system. By using various amino acids (X) in a cleavable ubiquitin-X-A3G fusion, we demonstrate that the half-life (t1/2) of X-A3G can be manipulated. We show that A3G molecules with a half-life of 13 min are incorporated into virions, whereas those with a half-life shorter than 5 min were not. The amount of X-A3G incorporated into virions increases from 13 min (Phe-A3G) to 85 min (Asn-A3G) and remains constant after this time period. Interestingly, despite the presence of similar levels of Arg-A3G (t1/2 = 28 min) and Asp-A3G (t1/2 = 65 min) into HIV-1 DELTAvif virions, inhibition of viral infectivity was only evident in the presence of A3G proteins with a longer half-life (t1/2 >= 65 min).

Authors:
;  [1]; ; ;  [2];  [1]
  1. URIA-Centro Patogenese Molecular, Faculdade de Farmacia da Universidade Lisboa, Av. Das Forcas Armadas, Lisboa 1649-059 (Portugal)
  2. Department of Pathology and the Lautenberg Center for General and Tumor Immunology, Hebrew University, Hadassah Medical School, Jerusalem 91120 (Israel)
Publication Date:
OSTI Identifier:
21357560
Resource Type:
Journal Article
Journal Name:
Virology
Additional Journal Information:
Journal Volume: 393; Journal Issue: 2; Other Information: DOI: 10.1016/j.virol.2009.07.031; PII: S0042-6822(09)00468-1; Copyright (c) 2009 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Journal ID: ISSN 0042-6822
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; AIDS VIRUS; AMINO ACIDS; HALF-LIFE; INFECTIVITY; INHIBITION; PROTEINS; CARBOXYLIC ACIDS; MICROORGANISMS; ORGANIC ACIDS; ORGANIC COMPOUNDS; PARASITES; VIRUSES

Citation Formats

Cadima-Couto, Iris, Freitas-Vieira, Acilino, Instituto de Medicina Molecular, Lisboa, Nowarski, Roni, Britan-Rosich, Elena, Kotler, Moshe, Goncalves, Joao, and Instituto de Medicina Molecular, Lisboa. Ubiquitin-fusion as a strategy to modulate protein half-life: A3G antiviral activity revisited. United States: N. p., 2009. Web. doi:10.1016/j.virol.2009.07.031.
Cadima-Couto, Iris, Freitas-Vieira, Acilino, Instituto de Medicina Molecular, Lisboa, Nowarski, Roni, Britan-Rosich, Elena, Kotler, Moshe, Goncalves, Joao, & Instituto de Medicina Molecular, Lisboa. Ubiquitin-fusion as a strategy to modulate protein half-life: A3G antiviral activity revisited. United States. https://doi.org/10.1016/j.virol.2009.07.031
Cadima-Couto, Iris, Freitas-Vieira, Acilino, Instituto de Medicina Molecular, Lisboa, Nowarski, Roni, Britan-Rosich, Elena, Kotler, Moshe, Goncalves, Joao, and Instituto de Medicina Molecular, Lisboa. 2009. "Ubiquitin-fusion as a strategy to modulate protein half-life: A3G antiviral activity revisited". United States. https://doi.org/10.1016/j.virol.2009.07.031.
@article{osti_21357560,
title = {Ubiquitin-fusion as a strategy to modulate protein half-life: A3G antiviral activity revisited},
author = {Cadima-Couto, Iris and Freitas-Vieira, Acilino and Instituto de Medicina Molecular, Lisboa and Nowarski, Roni and Britan-Rosich, Elena and Kotler, Moshe and Goncalves, Joao and Instituto de Medicina Molecular, Lisboa},
abstractNote = {The human APOBEC3G (A3G) is a potent inhibitor of HIV-1 replication and its activity is suppressed by HIV-1 virion infectivity factor (Vif). Vif neutralizes A3G mainly by inducing its degradation in the proteasome and blocking its incorporation into HIV-1 virions. Assessing the time needed for A3G incorporation into virions is, therefore, important to determine how quickly Vif must act to induce its degradation. We show that modelling the intracellular half-life of A3G can induce its Vif-independent targeting to the ubiquitin-proteasome system. By using various amino acids (X) in a cleavable ubiquitin-X-A3G fusion, we demonstrate that the half-life (t1/2) of X-A3G can be manipulated. We show that A3G molecules with a half-life of 13 min are incorporated into virions, whereas those with a half-life shorter than 5 min were not. The amount of X-A3G incorporated into virions increases from 13 min (Phe-A3G) to 85 min (Asn-A3G) and remains constant after this time period. Interestingly, despite the presence of similar levels of Arg-A3G (t1/2 = 28 min) and Asp-A3G (t1/2 = 65 min) into HIV-1 DELTAvif virions, inhibition of viral infectivity was only evident in the presence of A3G proteins with a longer half-life (t1/2 >= 65 min).},
doi = {10.1016/j.virol.2009.07.031},
url = {https://www.osti.gov/biblio/21357560}, journal = {Virology},
issn = {0042-6822},
number = 2,
volume = 393,
place = {United States},
year = {Sun Oct 25 00:00:00 EDT 2009},
month = {Sun Oct 25 00:00:00 EDT 2009}
}