Capsid-CPSF6 Interaction Licenses Nuclear HIV-1 Trafficking to Sites of Viral DNA Integration

Achuthan, Vasudevan; Perreira, Jill M.; Sowd, Gregory A.; Puray-Chavez, Maritza; McDougall, William M.; Paulucci-Holthauzen, Adriana; Wu, Xiaolin; Fadel, Hind J.; Poeschla, Eric M.; Multani, Asha S.; Hughes, Stephen H.; Sarafianos, Stefan G.; Brass, Abraham L.; Engelman, Alan

doi:10.1016/j.chom.2018.08.002

Cited by 172 publications

(237 citation statements)

References 76 publications

(202 reference statements)

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“…Our results are consistent with the finding that wild type capsids do not become IFN-hypersensitive in CPSF6 knockout cells (Bulli et al, 2016). Therefore, our data support a model in which CPSF6 binding plays a role in integration targeting (Achuthan et al, 2018; Bejarano et al, 2019; Rasheedi et al, 2016; Schaller et al, 2011; Sowd et al, 2016) but it does not shield HIV-1 capsids from capsid-targeting restrictions. In contrast, loss of CypA binding sensitizes HIV-1 capsids to TRIM5alpha restriction (Kyusik’s paper).…”

Section: Discussionsupporting

confidence: 76%

TRIM34 acts with TRIM5 to restrict HIV and SIV capsids

OhAinle

Kim

Keceli

et al. 2019

Preprint

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1The HIV-1 capsid protein makes up the core of the virion and plays a critical role in early 2 steps of HIV replication. Due to its exposure in the cytoplasm after entry, HIV capsid is a 3 target for host cell factors that act directly to block infection such as TRIM5 and MxB. 4 Several host proteins also play a role in facilitating infection, including in the protection 5 of HIV-1 capsid from recognition by host cell restriction factors. Through an unbiased 6 screening approach, called HIV-CRISPR, we show that the Cyclophilin A-binding 7 deficient P90A HIV-1 capsid mutant becomes highly-sensitized to TRIM5alpha restriction 8 in IFN-treated cells. Further, the CPSF6-binding deficient, N74D HIV-1 capsid mutant is 9 sensitive to restriction mediated by human TRIM34, a close paralog of the well-10 characterized HIV restriction factor TRIM5. This restriction occurs at the step of reverse 11 transcription, is independent of interferon stimulation and limits HIV-1 infection in key 12 target cells of HIV infection including CD4+ T cells and monocyte-derived dendritic cells. 13 TRIM34 restriction requires TRIM5alpha as knockout or knockdown of TRIM5alpha 14 results in a loss of antiviral activity. TRIM34 can also restrict some SIV capsids. Through 15 immunofluorescence studies, we show that TRIM34 and TRIM5alpha colocalize to 16 cytoplasmic bodies and are more frequently observed to be associated with infecting 17 N74D capsids than with WT capsids. Our results identify TRIM34 as an HIV-1 CA-18 targeting restriction factor and highlight the potential role for heteromultimeric TRIM 19 interactions in contributing restriction of HIV-1 infection in human cells.

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Section: Discussionsupporting

confidence: 76%

TRIM34 acts with TRIM5 to restrict HIV and SIV capsids

OhAinle

Kim

Keceli

et al. 2019

Preprint

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“…As an example analyte, we used an Alexa Fluor 488-labelled peptide derived from CPSF6 (residues 313-327), a host cell protein that promotes nuclear import and integration targeting of the viral DNA via interactions with the CA lattice. 34,35 CPSF6 313-327 binds into a pocket formed between adjacent CA molecules in a hexamer ( Figure 4D) with low affinity (K D of 50-100 μM). 34,35 The analyte was injected into the flow channel at concentration (250 nM) that was >100-fold below the K D , i.e.…”

Section: Growth Of Fluorescent Capsid Tubes On the Sensor Surface Asmentioning

confidence: 99%

“…34,35 CPSF6 313-327 binds into a pocket formed between adjacent CA molecules in a hexamer ( Figure 4D) with low affinity (K D of 50-100 μM). 34,35 The analyte was injected into the flow channel at concentration (250 nM) that was >100-fold below the K D , i.e. conditions that should lead to less than 1% occupation of available binding sites.…”

Section: Growth Of Fluorescent Capsid Tubes On the Sensor Surface Asmentioning

confidence: 99%

Fluorescence biosensor for real-time interaction dynamics of host proteins with HIV-1 capsid tubes

Lau

Walsh

Wang

et al. 2019

Preprint

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The human immunodeficiency virus 1 (HIV-1) capsid serves as a binding platform for proteins and small molecules from the host cell that regulate various steps in the virus life cycle. However, there are currently no quantitative methods that use assembled capsid lattices for measuring host-pathogen interaction dynamics. Here we developed a single molecule fluorescence biosensor using self-assembled capsid tubes as biorecognition elements and imaged capsid binders using total internal reflection fluorescence microscopy in a microfluidic setup. The method is highly sensitive in its ability to observe and quantify binding, obtain dissociation constants, extract kinetics with an extended application of using more complex analytes that can accelerate characterisation of novel capsid binders.

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“…Host cellular proteins are known to play critical roles in HIV-1 integration site selection. For example, LEDGF/p75 and CPSF6 promote integration into actively transcribing genes residing in genedense regions 29,30,32,57,[59][60][61][62][63] . Here we showed that host cellular A3 expression shifts the accumulation of HIV-1 integration sites away from genes and towards SINEs in a dose-dependent manner.…”

Section: Discussionmentioning

confidence: 99%

Antiretroviral APOBEC3 Cytidine Deaminases Alter HIV-1 Provirus Integration Site Profiles

Ajoge

Renner

Kohio

et al. 2019

Preprint

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APOBEC3 (A3) proteins are host-encoded deoxycytidine deaminases that provide an innate immune barrier to retroviral infection, notably against HIV-1. Low levels of deamination are believed to contribute to the rapid genetic evolution of HIV-1, while the catalytic activity of these proteins can induce catastrophic hypermutation in proviral DNA leading to near-total HIV-1 restriction. So far, little is known about how A3 cytosine deaminases might impact HIV-1 proviral DNA integrations into human chromosomal DNA. Using a deep sequencing approach, we analyzed the influence of catalytic active and inactive APOBEC3F and APOBEC3G on HIV-1 integration site selections. DNA editing was detected at the extremities of the long terminal repeat regions of the virus. Both catalytic active and non-catalytic A3 mutants decreased insertions into gene coding sequences and increased integration sites into SINE elements, oncogenes and transcriptionsilencing non-B DNA features. Our data implicates A3 as a host factor influencing HIV-1 integration site selection and promotes a more transcriptionally silent integration site profile. Ajoge et al., 2019 3 GRAPHICAL ABSTRACT Schematic depicting the influence of APOBEC3 (A3) proteins on HIV integration site targeting. Left, in the absence of A3, HIV has a strong preference for integrating into genes. Right, both catalytic active and non-catalytic A3 mutants decrease integration into genes and increase integration into SINE elements and in transcription-silencing non-B DNA features.

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Capsid-CPSF6 Interaction Licenses Nuclear HIV-1 Trafficking to Sites of Viral DNA Integration

Cited by 172 publications

References 76 publications

TRIM34 acts with TRIM5 to restrict HIV and SIV capsids

TRIM34 acts with TRIM5 to restrict HIV and SIV capsids

Fluorescence biosensor for real-time interaction dynamics of host proteins with HIV-1 capsid tubes

Antiretroviral APOBEC3 Cytidine Deaminases Alter HIV-1 Provirus Integration Site Profiles

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