Capsid-Dependent Host Factors in HIV-1 Infection

Yamashita, Masahiro; Engelman, Alan

doi:10.1016/j.tim.2017.04.004

Cited by 110 publications

(120 citation statements)

References 149 publications

(355 reference statements)

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“…29 The experiments presented here demonstrate that, in primary human blood cells, 30 HIV-1 exploits CypA to evade CA recognition by, and the antiviral activity of, 31 endogenous TRIM5α. This answers the long-standing question of how CypA promotes 1 HIV-1 infection and clearly establishes that, in the absence of CypA, human TRIM5α 2 potently restricts HIV-1. Conservation of the lentiviral CA-CypA interaction across 3 millions of years of evolution is likely a result of selective pressure applied by TRIM5α 4 orthologues encoded by host species that are otherwise permissive for lentiviral 5 replication.…”

supporting

confidence: 60%

“…2b, c). As compared 1 to the wild-type, the infectivity of vector bearing CA-P90A was decreased ( Fig. 2b, c), 2 and the infectivity of this mutant was rescued by TRIM5 shRNA (Fig.…”

mentioning

confidence: 91%

“…3b). Either of two non-immunosuppressive CypA inhibitors 1 derived from sanglifehrin A 37 , decreased HIV-1 transduction efficiency in primary CD4 + 2 T cells ( Fig. 1f; Extended Data Fig.…”

mentioning

confidence: 99%

“…28 To determine if TRIM5α associates with HIV-1 CA in cells when the CA-CypA 29 interaction is disrupted, primary human macrophages were stably transduced with 30 TRIM5 shRNA or Luc shRNA and then challenged for 2 hrs with wild-type HIV-1 reporter vector, in the presence or absence of CsA. Cells were fixed and subjected to 1 the proximity ligation assay (PLA) with antibodies specific for HIV-1 CA and 2 endogenous human TRIM5α. When cells were challenged with HIV-1 in the absence of 3 CsA, very few puncta were detected ( Fig.…”

mentioning

confidence: 99%

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Cyclophilin A protects HIV-1 from restriction by human TRIM5α

Kim

Dauphin

Kömürlü

et al. 2019

Preprint

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The capsid (CA) protein lattice of HIV-1 and other retroviruses encases viral 1 genomic RNA and regulates steps that are essential to retroviral invasion of 2 target cells, including reverse transcription, nuclear trafficking, and integration of 3 viral cDNA into host chromosomal DNA 1 . Cyclophilin A (CypA), the first cellular 4 protein reported to bind HIV-1 CA 2 , has interacted with invading lentiviruses 5 related to HIV-1 for millions of years 3-7 . Disruption of the CA-CypA interaction 6 decreases HIV-1 infectivity in human cells 8-12 , but stimulates infectivity in non-7 human primate cells 13-15 . Genetic and biochemical data suggest that CypA 8 interaction with CA protects HIV-1 from a restriction factor in human cells 16-20 . 9 Discovery of the CA-specific restriction factor TRIM5α 21 , and of TRIM5-CypA 10 fusion genes that were independently generated at least four times in 11 phylogeny 4,5,15,22-25 , pointed to human TRIM5α as the CypA-sensitive restriction 12 factor. However, significant HIV-1 restriction by human TRIM5α 21 , let alone 13 inhibition of such activity by CypA 26 , has not been detected. Here, exploiting 14 reverse genetic tools optimized for primary human CD4 + T cells, macrophages, 15 and dendritic cells, we demonstrate that disruption of the CA-CypA interaction 16 renders HIV-1 susceptible to restriction by human TRIM5α, with the block 17 occurring before reverse transcription. Identical findings were obtained with 18 single-cycle vectors or with replication-competent HIV-1, including sexually-19 transmitted clones from sub-Saharan Africa. Endogenous TRIM5α was observed 20 to associate with virion cores as they entered the macrophage cytoplasm, but 21 only when the CA-CypA interaction was disrupted. These experiments resolve the 22 long-standing mystery of the role of CypA in HIV-1 replication by demonstrating 23 that this ubiquitous cellular protein shields HIV-1 from previously inapparent, but 24 potent inhibition, imposed by human TRIM5α. Hopefully this reinvigorates 25 development of CypA-inhibitors for treatment of HIV-1 and other CypA-dependent 26 pathogens 27-30 . 27To assess the role of TRIM5α and CypA in the primary human blood cell types 1 that serve as targets for HIV-1 infection in vivo, lentiviral vectors were optimized for titer 2 and knockdown efficiency in these cells 26,[31][32][33][34] . Human macrophages, dendritic cells, 3 and CD4 + T cells were transduced with lentivectors bearing a puromycin resistance 4 cassette and shRNAs targeting either TRIM5 or luciferase (Luc) as a control. After three 5 days of selection in puromycin, knockdown was confirmed by RT-qPCR for TRIM5 6 mRNA, and by rescue of N-MLV restriction (Extended Data Fig. 2a-c), as done 7 previously 26,31 . TRIM5 and Luc control knockdown cells were then challenged with 8 single-cycle, VSV G-pseudotyped, HIV-1-GFP reporter vectors. Three days later, the 9 percentage of GFP + cells was assessed by flow cytometry as a measure of infectivity 10

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supporting

confidence: 60%

“…2b, c). As compared 1 to the wild-type, the infectivity of vector bearing CA-P90A was decreased ( Fig. 2b, c), 2 and the infectivity of this mutant was rescued by TRIM5 shRNA (Fig.…”

mentioning

confidence: 91%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Cyclophilin A protects HIV-1 from restriction by human TRIM5α

Kim

Dauphin

Kömürlü

et al. 2019

Preprint

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“…We mainly introduced mutations into helical domains. It is already known that the loop structure between helices 4 and 5 (4 -5 loop) serves as binding site for cellular proline isomerase cyclophilin A, regulating viral replication (2,7,8). The 4-5 and 6-7 loops in CA-NTD are shown to be critical by us for human specific nature of HIV-1 as viral determinants for narrow HIV-1 species tropism (16,23).…”

Section: Generation Of Hiv-1 Ca-ntd Mutantsmentioning

confidence: 99%

Virological characterization of HIV‐1 CA‐NTD mutants constructed in a virus‐lineage reflected manner

et al. 2018

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: Capsid (CA) protein is a major virion-constituent of all retroviruses including human immunodeficiency virus type 1 (HIV-1), and is essential for early and late phases in viral replication cycle through interaction with numerous cellular factors. In particular, N-terminal domain (NTD) of HIV-1 CA has been frequently and well reported to bind to various host cell proteins that considerably affect viral replication potential. In this study, in order to better define biological bases of the CA-NTD for HIV-1 replication, we performed an extensive mutational analysis in an unprecedented manner. By aligning CA-NTD sequences derived from representative infectious molecular clones of HIV-1, HIV-2, and simian immunodeficiency virus isolated from the rhesus macaque (SIVmac), a number of amino acids specific to HIV-1 were selected, and were replaced with those from SIVmac at the corresponding sites. Mutant viruses thus generated were then examined for multi -cycle infectivity, single-cycle infectivity, and ability to produce progeny virions. While some CA-NTD mutations affected viral replication ability to varying degrees, those in helix 7 abolished viral growth potential without exception. These results highlight functional importance of non -conserved amino acids in helix 7, and give new insights into functionality of HIV-1 CA-NTD.

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Cellular and molecular mechanisms of HIV-1 integration targeting

Engelman

Singh

2018

Cell. Mol. Life Sci.

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Integration is central to HIV-1 replication and helps mold the reservoir of cells that persists in AIDS patients. HIV-1 interacts with specific cellular factors to target integration to interior regions of transcriptionally active genes within gene-dense regions of chromatin. The viral capsid interacts with several proteins that are additionally implicated in virus nuclear import, including cleavage and polyadenylation specificity factor 6, to suppress integration into heterochromatin. The viral integrase protein interacts with transcriptional co-activator lens epithelium-derived growth factor p75 to principally position integration within gene bodies. The integrase additionally senses target DNA distortion and nucleotide sequence to help fine-tune the specific phosphodiester bonds that are cleaved at integration sites. Research into virus-host interactions that underlie HIV-1 integration targeting has aided the development of a novel class of integrase inhibitors and may help to improve the safety of viral-based gene therapy vectors.

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Capsid-Dependent Host Factors in HIV-1 Infection

Cited by 110 publications

References 149 publications

Cyclophilin A protects HIV-1 from restriction by human TRIM5α

Cyclophilin A protects HIV-1 from restriction by human TRIM5α

Virological characterization of HIV‐1 CA‐NTD mutants constructed in a virus‐lineage reflected manner

Cellular and molecular mechanisms of HIV-1 integration targeting

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