A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression

ABSTRACT Extensive genetic diversity is a defining characteristic of human immunodeficiency virus type 1 (HIV-1) and poses a significant barrier to the development of an effective vaccine. To better understand the impact of this genetic diversity on the HIV-1 pathogenic factor Nef, we compiled a pan...

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Autores principales:	Aaron L. Johnson, Brennan S. Dirk, Mathieu Coutu, S. M. Mansour Haeryfar, Eric J. Arts, Andrés Finzi, Jimmy D. Dikeakos
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Nef diversity human immunodeficiency virus Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/f2e1efd322d84669aeb8e11a593106fa
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spelling	oai:doaj.org-article:f2e1efd322d84669aeb8e11a593106fa2021-11-15T15:22:03ZA Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression10.1128/mSphere.00288-162379-5042https://doaj.org/article/f2e1efd322d84669aeb8e11a593106fa2016-12-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00288-16https://doaj.org/toc/2379-5042ABSTRACT Extensive genetic diversity is a defining characteristic of human immunodeficiency virus type 1 (HIV-1) and poses a significant barrier to the development of an effective vaccine. To better understand the impact of this genetic diversity on the HIV-1 pathogenic factor Nef, we compiled a panel of reference strains from the NIH Los Alamos HIV Database. Initial sequence analysis identified point mutations at Nef residues 13, 84, and 92 in subtype C reference strain C.BR92025 from Brazil. Functional analysis revealed impaired major histocompatibility complex class I and CD4 downregulation of strain C.BR92025 Nef, which corresponded to decreased protein expression. Metabolic labeling demonstrated that strain C.BR92025 Nef has a greater rate of protein turnover than subtype B reference strain B.JRFL that, on the basis of mutational analysis, is related to Nef residue A84. An alanine-to-valine substitution at position 84, located in alpha helix 2 of Nef, was sufficient to alter the rate of turnover of an otherwise highly expressed Nef protein. In conclusion, these findings highlight HIV-1 Nef residue A84 as a major determinant of protein expression that may offer an additional avenue to disrupt or mediate the effects of this key HIV-1 pathogenic factor. IMPORTANCE The HIV-1 Nef protein has been established as a key pathogenic determinant of HIV/AIDS, but there is little knowledge of how the extensive genetic diversity of HIV-1 affects Nef function. Upon compiling a set of subtype-specific reference strains, we identified a subtype C reference strain, C.BR92025, that contained natural polymorphisms at otherwise highly conserved residues 13, 84, and 92. Interestingly, strain C.BR92025 Nef displayed impaired Nef function and had decreased protein expression. We have demonstrated that strain C.BR92025 Nef has a higher rate of protein turnover than highly expressed Nef proteins and that this higher rate of protein turnover is due to an alanine-to-valine substitution at Nef residue 84. These findings highlight residue A84 as a major determinant of HIV-1 Nef expression.Aaron L. JohnsonBrennan S. DirkMathieu CoutuS. M. Mansour HaeryfarEric J. ArtsAndrés FinziJimmy D. DikeakosAmerican Society for MicrobiologyarticleNefdiversityhuman immunodeficiency virusMicrobiologyQR1-502ENmSphere, Vol 1, Iss 6 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Nef diversity human immunodeficiency virus Microbiology QR1-502
spellingShingle	Nef diversity human immunodeficiency virus Microbiology QR1-502 Aaron L. Johnson Brennan S. Dirk Mathieu Coutu S. M. Mansour Haeryfar Eric J. Arts Andrés Finzi Jimmy D. Dikeakos A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression
description	ABSTRACT Extensive genetic diversity is a defining characteristic of human immunodeficiency virus type 1 (HIV-1) and poses a significant barrier to the development of an effective vaccine. To better understand the impact of this genetic diversity on the HIV-1 pathogenic factor Nef, we compiled a panel of reference strains from the NIH Los Alamos HIV Database. Initial sequence analysis identified point mutations at Nef residues 13, 84, and 92 in subtype C reference strain C.BR92025 from Brazil. Functional analysis revealed impaired major histocompatibility complex class I and CD4 downregulation of strain C.BR92025 Nef, which corresponded to decreased protein expression. Metabolic labeling demonstrated that strain C.BR92025 Nef has a greater rate of protein turnover than subtype B reference strain B.JRFL that, on the basis of mutational analysis, is related to Nef residue A84. An alanine-to-valine substitution at position 84, located in alpha helix 2 of Nef, was sufficient to alter the rate of turnover of an otherwise highly expressed Nef protein. In conclusion, these findings highlight HIV-1 Nef residue A84 as a major determinant of protein expression that may offer an additional avenue to disrupt or mediate the effects of this key HIV-1 pathogenic factor. IMPORTANCE The HIV-1 Nef protein has been established as a key pathogenic determinant of HIV/AIDS, but there is little knowledge of how the extensive genetic diversity of HIV-1 affects Nef function. Upon compiling a set of subtype-specific reference strains, we identified a subtype C reference strain, C.BR92025, that contained natural polymorphisms at otherwise highly conserved residues 13, 84, and 92. Interestingly, strain C.BR92025 Nef displayed impaired Nef function and had decreased protein expression. We have demonstrated that strain C.BR92025 Nef has a higher rate of protein turnover than highly expressed Nef proteins and that this higher rate of protein turnover is due to an alanine-to-valine substitution at Nef residue 84. These findings highlight residue A84 as a major determinant of HIV-1 Nef expression.
format	article
author	Aaron L. Johnson Brennan S. Dirk Mathieu Coutu S. M. Mansour Haeryfar Eric J. Arts Andrés Finzi Jimmy D. Dikeakos
author_facet	Aaron L. Johnson Brennan S. Dirk Mathieu Coutu S. M. Mansour Haeryfar Eric J. Arts Andrés Finzi Jimmy D. Dikeakos
author_sort	Aaron L. Johnson
title	A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression
title_short	A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression
title_full	A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression
title_fullStr	A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression
title_full_unstemmed	A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression
title_sort	highly conserved residue in hiv-1 nef alpha helix 2 modulates protein expression
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/f2e1efd322d84669aeb8e11a593106fa
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A Highly Conserved Residue in HIV-1 Nef Alpha Helix 2 Modulates Protein Expression

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