Mutagenic analysis of human immunodeficiency virus type 1 Vpr: role of a predicted N-terminal alpha-helical structure in Vpr nuclear localization and virion incorporation

Yao, Xiaojian; Subbramanian, Ramu A.; Rougeau, Nicole; Boisvert, Françoise; Bergeron, Dominique; Cohen, Éric A.

doi:10.1128/jvi.69.11.7032-7044.1995

Cited by 166 publications

(124 citation statements)

References 46 publications

(68 reference statements)

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“…Mutation of Ala30 to leucine completely abolishes G 2 cell cycle arrest and virion packaging of Vpr [23]. Mutations of either Leu23 or Ala30 to phenylalanine dramatically reduce virion incorporation of Vpr but do not interfere with its nuclear localization [24]. One may speculate, that the leucinezipper like sequence motif preceding Ala30 serves together with helical secondary structure as a basis for a variety of protein±protein interactions between Vpr and other viral and cellular factors.…”

Section: Discussionmentioning

confidence: 99%

Solution structure of human immunodeficiency virus type 1 Vpr(13-33) peptide in micelles

Engler¹,

Stangler²,

Willbold³

2001

Eur J Biochem

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Human immunodeficiency virus type 1 protein R (HIV-1 Vpr) promotes nuclear entry of viral nucleic acids in nondividing cells, causes G 2 cell cycle arrest and is involved in cellular differentiation and cell death. Also, Vpr subcellular localization is as variable as its functions. It is known that, consistent with its role in nuclear transport, Vpr localizes to the nuclear envelope of human cells. Further, a reported ion channel activity of Vpr obviously is dependent on its localization in or at membranes. We focused our structural studies on the secondary structure of a peptide consisting of residues 13±33 of HIV-1 Vpr in micelles. Employing nuclear magnetic resonance and circular dichroism spectroscopy we found this part of Vpr, known to be essential for nuclear localization, to be almost completely a helical. Our results provide structural data suggesting residues 13±33 of Vpr to form an amphipathic, leucine-zipper-like a helix that serves as a basis for interactions with a variety of viral and cellular factors.Keywords: HIV-1; Vpr; peptide solution structure; dodecylphosphocholine; micelles.Human immunodeficiency virus type 1 (HIV-1) is a member of the lentivirus family. In addition to the gag, pol and env genes present in all retroviruses, HIV-1 encodes regulatory and accessory proteins, that are decisive for viral infectivity, replication and pathogenity. One of these proteins is virus protein R (Vpr). Vpr seems to be required at various steps of the HIV replication cycle and is therefore an interesting target for the development of antiviral agents. This 96-amino-acid protein is an important factor for the pathogenicy of HIV [1,2]. Vpr is an integral part of viral particles suggesting an important role in early stages of infection [3±6]. Furthermore, Vpr is involved in the transport of the preintegration complex into the host cell nucleus, which is an important feature for infection of nondividing cells [7,8] Recent structural studies of Vpr fragments by NMR were performed in trifluoroethanol (30%) containing solution and, not surprisingly, revealed a long amphipathic a helixturn-a helix (amino acids 17±46) motif ended by a turn for Vpr (1±51) [20]. The Vpr (52±96) fragment, also in trifluorethanol containing solution, is characterized by an amphipathic a helix from residue 53 to residue 78 and a less defined C-terminal domain [21]. Another fragment of Vpr (50±82) was shown to contain a helix from residues 53±81 in 50% trifluorethanol. Trifluorethanol, however, is well known to induce a helical secondary structures in peptides [22].We focused our structural studies on residues 13±33 of HIV-1 Vpr. This part of Vpr is known to be essential for ion chanel activity [15], cell cycle arrest [23], incorporation of Vpr into virus-like particles [24±26], and its nuclear localization [27]. Because a membranous or membranelike environment may be important for most of these activities, we carried out structural studies of Vpr(13±33) peptide in dodecylphosphocholine (dodecyl-PCho) containing micelles by CD and NMR sp...

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Section: Discussionmentioning

confidence: 99%

Solution structure of human immunodeficiency virus type 1 Vpr(13-33) peptide in micelles

Engler¹,

Stangler²,

Willbold³

2001

Eur J Biochem

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“…The various isogenic proviral type or mutant Env and the G glycoprotein of VSV. In order to ensure that VSV-G is incorporated in HIV virions even in the presence of Env, constructs were obtained as follows: vprϩ/-: HxBRU and HxBRURwith a stop codon at the initiation site of the vpr gene of HIV-BRU 10 6 COS cells were transfected with 10 μg of either proviral construct with or without 10 μg of SVCMV-VSV-G by the lipofection method have been described elsewhere (Lavallée et al, 1994;Yao et al, 1995); vpuϩ/-: HXBH10 and HXBH10vpu-, were described previously (Yao (Leventis and Silvius, 1990;Mouland et al, 1994). Cells were labeled 40 h after transfection with 150 μCi/ml of [ 35 S]methionine for 8 h. et al, 1992); nefϩ/-: pNL4.3-based pNLV102 and pNLV102ΔXho, gifts of Dr S.Venkatesan, and described previously (Ahmad and Venkatesan, Supernatants were harvested and ultracentrifuged (30 000 r.p.m.…”

Section: Incorporation Of Vsv-g In Hiv Virionsmentioning

confidence: 99%

The membrane-proximal intracytoplasmic tyrosine residue of HIV-1 envelope glycoprotein is critical for basolateral targeting of viral budding in MDCK cells

et al. 1997

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Budding of retroviruses from polarized epithelial Madin–Darby canine kidney cells (MDCK) takes place specifically at the basolateral membrane surface. This sorting event is suspected to require a specific signal harbored by the viral envelope glycoprotein and it was previously shown that, as for most basolateral proteins, the intracytoplasmic domain plays a crucial role in this targeting phenomenon. It is well known that tyrosine‐based motifs are a central element in basolateral targeting signals. In the present study, site‐directed mutagenesis was used to generate conservative or non‐conservative substitutions of each four intracytoplasmic tyrosines of the human immunodeficiency virus (HIV‐1) envelope glycoprotein. This approach revealed that the membrane‐proximal tyrosine is essential to ensure both the basolateral localization of envelope glycoprotein and the basolateral targeting of HIV‐1 virions. Substitutions of the membrane‐proximal tyrosine did not appear to affect incorporation of envelope glycoprotein into the virions, as assayed by virion infectivity and protein content, nor its capability to ensure its role in viral infection, as determined by viral multiplication kinetics. Altogether, these results indicate that the intracytoplasmic domain of the HIV‐1 envelope glycoprotein harbors a unique, tyrosine‐based, basolateral targeting signal. Such a tyrosine‐based targeting signal may play a fundamental role in HIV transmission and pathogenesis.

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“…Although numerous studies have demonstrated that HIV‐1 Vpr mutations could dramatically affect its known functions , these mutations were generated in the lab and did not represent the profile of the naturally occurring mutations over the course of infection. Indeed, in order to address the correlation between the structural domains and Vpr function, a set of molecular strategies involving deletion, insertion and/or substitution mutagenesis was developed.…”

Section: Vpr Mutations‐dependent Phenotypesmentioning

confidence: 99%

“…Nevertheless, it should also be considered that it may be easy to disrupt the interactions of Vpr by disrupting its structure, because it is a relatively small protein. The tertiary structure of Vpr was determined by Morellet and colleagues (2003), who performed a nuclear magnetic resonance analysis of the full‐length polypeptide . Vpr contains three α‐helical domains with amphipathic properties, linked by loops and folded around a hydrophobic core surrounded by a flexible N‐terminal domain and a C‐terminal arginine‐rich region that are negatively and positively charged, respectively (Figure ).…”

Section: Introductionmentioning

confidence: 99%

HIV1‐viral protein R (Vpr) mutations: associated phenotypes and relevance for clinical pathologies

Soares

Rocha

Meliço-Silvestre

et al. 2016

Reviews in Medical Virology

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Over the last 30 years, research into HIV has advanced the knowledge of virus genetics and the development of efficient therapeutic strategies. HIV-1 viral protein R (Vpr) is a specialized and multifunctional protein that plays important roles at multiple stages of the HIV-1 viral life cycle. This protein interacts with a number of cellular and viral proteins and with multiple activities including nuclear transport of the pre-integration complex (PIC) to the nucleus, transcriptional activation, cell cycle arrest at G2/M transition phase and induction of cell death via apoptosis. Specifically, Vpr has been shown to control many host cell functions through a variety of biological processes and by interaction with several cellular pathways. The different functions of Vpr may enhance viral replication and impair the immune system in HIV-1 infected patients. Importantly, functional defects induced by mutations in the Vpr protein correlate with slow disease progression of HIV-infected patients. Vpr is also associated with other concomitant pathologies developed by these patients, which may lead it to be considered as a potential novel therapeutic target. This review will focus on HIV-1 Vpr, mainly on the importance of its structural mutations on the progression of HIV infection, associated phenotypes and relevance for clinical pathologies. Copyright © 2016 John Wiley & Sons, Ltd.

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Mutagenic analysis of human immunodeficiency virus type 1 Vpr: role of a predicted N-terminal alpha-helical structure in Vpr nuclear localization and virion incorporation

Cited by 166 publications

References 46 publications

Solution structure of human immunodeficiency virus type 1 Vpr(13-33) peptide in micelles

Solution structure of human immunodeficiency virus type 1 Vpr(13-33) peptide in micelles

The membrane-proximal intracytoplasmic tyrosine residue of HIV-1 envelope glycoprotein is critical for basolateral targeting of viral budding in MDCK cells

HIV1‐viral protein R (Vpr) mutations: associated phenotypes and relevance for clinical pathologies

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