Novel mutant human immunodeficiency virus type 1 strains with high degree of resistance to cynomolgus macaque TRIMCyp generated by random mutagenesis

Sultana, Tahmina; Nakayama, Emi E.; Tobita, Satoshi; Yokoyama, Masahiro; Seki, Yusuke; Saito, Akatsuki; Nomaguchi, Masako; Adachi, Akio; Akari, Hirofumi; Sato, Hironori; Shioda, Tatsuo

doi:10.1099/jgv.0.000408

Cited by 9 publications

(15 citation statements)

References 38 publications

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“…We first examined the IFN-β sensitivity of the RGDA/Q112D virus (40), since the P90A mutation in CA, which results in a CypA binding-deficient mutant (42, 43), has been shown to increase the sensitivity to IFN-α-mediated inhibition (10). We used Jurkat cells, a T cell line, since T cells are the major target for HIV-1 replication.…”

Section: Resultsmentioning

confidence: 99%

“…To establish a macaque model for HIV-1 infection, we and others constructed HIV-1 derivatives capable of establishing productive infection in monkey cells by manipulating the CA sequence, a major determinant for species specificity (36–39). We recently reported that one mutant virus, called the RGDA/Q112D virus, which contains the H87R, A88G, P90D, P93A, and Q112D changes in CA, was highly resistant to cynomolgus monkey (CM) TRIMCyp (40). An interesting property of this RGDA/Q112D virus is its behavior in cells coinfected with Sendai virus (SeV).…”

Section: Introductionmentioning

confidence: 99%

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Multiple Pathways To Avoid Beta Interferon Sensitivity of HIV-1 by Mutations in Capsid

Sultana

Mamede

Saito

et al. 2019

J Virol

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HIV-1 infection causes robust innate immune activation in virus-infected patients. This immune activation is characterized by elevated levels of type I interferons (IFNs), which can block HIV-1 replication. Recent studies suggest that the viral capsid protein (CA) is a determinant for the sensitivity of HIV-1 to IFN-mediated restriction. Specifically, it was reported that the loss of CA interactions with CPSF6 or CypA leads to higher IFN sensitivity. However, the molecular mechanism of CA adaptation to IFN sensitivity is largely unknown. Here, we experimentally evolved an IFN-β-hypersensitive CA mutant which showed decreased binding to CPSF6 and CypA in IFN-β-treated cells. The CA mutations that emerged from this adaptation indeed conferred IFN-β resistance. Our genetic assays suggest a limited contribution of known host factors to IFN-β resistance. Strikingly, one of these mutations accelerated the kinetics of reverse transcription and uncoating. Our findings suggest that HIV-1 selected multiple, known host factor-independent pathways to avoid IFN-β-mediated restriction.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Multiple Pathways To Avoid Beta Interferon Sensitivity of HIV-1 by Mutations in Capsid

Sultana

Mamede

Saito

et al. 2019

J Virol

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“…Monolayer cell lines HEK293T (ATCC CRL-1573), HeLa (ATCC CCL-2), and HeLa-derived reporter TZM-bl (58) were cultured and maintained in Eagle's minimal essential medium containing 10% heat-Molecular modeling of HIV-1 capsid protein. HIV-1 capsid monomer structures with various linker and MHR mutations were constructed by the homology modeling method with Molecular Operating Environment (MOE; Chemical Computing Group Inc., Montreal, QC, Canada) as described previously for modeling of various HIV-1 capsid mutants (64,(68)(69)(70). The reported structures of HIV-1 immature capsid (PDB code 4USN) (25) and mature capsid (PDB code 3J34) (55) were used as templates for modeling the immature and mature CA proteins, respectively, of HIV-1 NL4-3 and its derivatives.…”

Section: Cellsmentioning

confidence: 99%

Allosteric Regulation of HIV-1 Capsid Structure for Gag Assembly, Virion Production, and Viral Infectivity by a Disordered Interdomain Linker

Koma

Kotani

Miyakawa

et al. 2019

J Virol

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The retroviral Gag capsid (Gag-CA) interdomain linker is an unstructured peptide segment connecting structured N-terminal and C-terminal domains. Although the region is reported to play roles in virion morphogenesis and infectivity, underlying molecular mechanisms remain unexplored. To address this issue, we determined biological and molecular phenotypes of HIV-1 CA linker mutants by experimental and in silico approaches. Among the nine linker mutants tested, eight exhibited attenuation of viral particle production to various extents mostly in parallel with a reduction in viral infectivity. Sucrose density gradient, confocal microscopy, and live-cell protein interaction analyses indicated that the defect is accompanied by attenuation of Gag-Gag interactions following Gag plasma membrane targeting in the cells. In silico analyses revealed distinct distributions of interaction-prone hydrophobic patches between immature and mature CA proteins. Molecular dynamics simulations predicted that the linker mutations can allosterically alter structural fluctuations, including the interaction surfaces apart from the mutation sites in both the immature and mature CA proteins. These results suggest that the HIV-1 CA interdomain linker is a cis-modulator of the CA interaction surfaces to optimize efficiency of Gag assembly, virion production, and viral infectivity. IMPORTANCE HIV-1 particle production and infection are highly ordered processes. Viral Gag proteins play a central role in the assembly and disassembly of viral molecules. Of these, capsid protein (CA) is a major contributor to the Gag-Gag interactions. CA consists of two structured domains, i.e., N-terminal (NTD) and C-terminal (CTD) domains, connected by an unstructured domain named the interdomain linker. While multiple regions in the NTD and CTD are reported to play roles in virion morphogenesis and infectivity, the roles of the linker region in Gag assembly and virus particle formation remain elusive. In this study, we showed by biological and molecular analyses that the linker region functions as an intramolecular modulator to tune Gag assembly, virion production, and viral infectivity. Our study thus illustrates a hitherto-unrecognized mechanism, an allosteric regulation of CA structure by the disordered protein element, for HIV-1 replication.

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“…When we examined infection in cells co-infected with an MxB-expressing SeV, the reporter virus with NL4-3 capsid (1L) was suppressed nearly 30-fold, as we expected. In contrast, a reporter virus in which the loop between α-helices 4 and 5 of the NL4-3 capsid was replaced with the corresponding loop of SIVmac 31 (2L) showed resistance to MxB-mediated restriction, since 2L showed only twofold change in viral titer in the presence of MxB ( Fig. 5 ).…”

Section: Resultsmentioning

confidence: 98%

Naturally Occurring Mutations in HIV-1 CRF01_AE Capsid Affect Viral Sensitivity to Restriction Factors

Nakayama

Saito

Sultana

et al. 2018

AIDS Research and Human Retroviruses

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TRIM5α and MxB are known as restriction factors that inhibit the early step of intracellular HIV-1 replication cycle. Both factors are believed to interact with the incoming virus core to suppress HIV-1 infection. The extreme diversity of HIV-1 is thought to be a consequence of its propensity to mutate to escape immune responses and host restriction factors. We recently determined the capsid sequences for 144 HIV-1 CRF01_AE viruses obtained in Thailand from 2005 to 2011. In this study, we further analyzed the amino acid variations among the capsid sequences of 204 HIV-1 CRF01_AE obtained in Thailand and China, including 84 of the aforementioned 144 viruses, to detect mutations permitting escape from restriction by host factors. We found a characteristic combination of E79D, V83T, and H87Q in sequences from Chinese viruses and subsequently showed that this combination conferred partial resistance to MxB. Interestingly, this combination conferred resistance to human TRIM5α as well. The H87Q mutation alone conferred resistance to MxB in the CRF01_AE background, but not in subtype B virus. In contrast, the H87Q mutation alone conferred resistance to human TRIM5α in both the CFR01_AE and subtype B backgrounds. BLAST analysis revealed the presence of the E79D, V83T, and H87Q combination in CRF01_AE viruses isolated not only in China but also in many other countries. Although the mechanistic details as well as precise role of MxB antiviral activity in infected individuals remain to be clarified, our data suggest an interaction between MxB and the HIV-1 capsid in vivo.

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Novel mutant human immunodeficiency virus type 1 strains with high degree of resistance to cynomolgus macaque TRIMCyp generated by random mutagenesis

Cited by 9 publications

References 38 publications

Multiple Pathways To Avoid Beta Interferon Sensitivity of HIV-1 by Mutations in Capsid

Multiple Pathways To Avoid Beta Interferon Sensitivity of HIV-1 by Mutations in Capsid

Allosteric Regulation of HIV-1 Capsid Structure for Gag Assembly, Virion Production, and Viral Infectivity by a Disordered Interdomain Linker

Naturally Occurring Mutations in HIV-1 CRF01_AE Capsid Affect Viral Sensitivity to Restriction Factors

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