Nucleoporin NUP153 Phenylalanine-Glycine Motifs Engage a Common Binding Pocket within the HIV-1 Capsid Protein to Mediate Lentiviral Infectivity

Matreyek, Kenneth A.; Yücel, Sara Suna; Li, Xiang; Engelman, Alan

doi:10.1371/journal.ppat.1003693

Cited by 249 publications

(399 citation statements)

References 90 publications

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“…HIV-1 dependence on TNPO3 and Nup153 is altered by substitutions in the PF74 binding site (13,40,47), suggesting that the compound competes with host factor binding. Studies in our laboratory and others also suggest that HIV-1 sensitivity to PF74 is dependent on the ability of the virus to engage these host proteins.…”

Section: Discussionmentioning

confidence: 99%

HIV-1 Resistance to the Capsid-Targeting Inhibitor PF74 Results in Altered Dependence on Host Factors Required for Virus Nuclear Entry

Zhou

Price

Halambage

et al. 2015

J Virol

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During HIV-1 infection of cells, the viral capsid plays critical roles in reverse transcription and nuclear entry of the virus. The capsid-targeting small molecule PF74 inhibits HIV-1 at early stages of infection. HIV-1 resistance to PF74 is complex, requiring multiple amino acid substitutions in the viral CA protein. Here we report the identification and analysis of a novel PF74-resistant mutant encoding amino acid changes in both domains of CA, three of which are near the pocket where PF74 binds. Interestingly, the mutant virus retained partial PF74 binding, and its replication was stimulated by the compound. The mutant capsid structure was not significantly perturbed by binding of PF74; rather, the mutations inhibited capsid interactions with CPSF6 and Nup153 and altered HIV-1 dependence on these host factors and on TNPO3. Moreover, the replication of the mutant virus was markedly impaired in activated primary CD4 ؉ T cells and macrophages. Our results suggest that HIV-1 escapes a capsid-targeting small molecule inhibitor by altering the virus's dependence on host factors normally required for entry into the nucleus. They further imply that clinical resistance to inhibitors targeting the PF74 binding pocket is likely to be strongly limited by functional constraints on HIV-1 evolution. IMPORTANCE The HIV-1 capsid plays critical roles in early steps of infection and is an attractive target for therapy. Here we show that selection for resistance to a capsid-targeting small molecule inhibitor can result in viral dependence on the compound. The mutant virus was debilitated in primary T cells and macrophages-cellular targets of infection in vivo.The mutations also altered the virus's dependence on cellular factors that are normally required for HIV-1 entry into the nucleus. This work provides new information regarding mechanisms of HIV-1 resistance that should be useful in efforts to develop clinically useful drugs targeting the HIV-1 capsid.T he HIV-1 capsid plays multiple roles during the early stages of infection, all of which are critical for optimal infectivity. The conical viral capsid, composed of the CA protein, consists of a lattice of hexamers interspersed with 12 pentamers, which provide curvature and closure of the ends of the cone (reviewed in reference 1). Studies of the viral capsid have received much attention of late, with the identification of inhibitory host proteins that directly target the viral capsid (2-8) and elucidation of high-resolution structures of the CA hexamer and pentamer and tubular assemblies of CA resulting in working structural models of the entire viral capsid (9-11). Several positive-acting host factors have been reported to bind the viral capsid, including cyclophilin A, CPSF6, TNPO3, Nup153, and RanBP2 (12-21); in general, these appear to facilitate infection by coordinating entry of the viral preintegration complex into the nucleus, where they may also play a role in chromatin targeting during integration. Moreover, the host factors CPSF6 and cyclophilin A contribut...

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Section: Discussionmentioning

confidence: 99%

HIV-1 Resistance to the Capsid-Targeting Inhibitor PF74 Results in Altered Dependence on Host Factors Required for Virus Nuclear Entry

Zhou

Price

Halambage

et al. 2015

J Virol

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“…Several genome-wide RNA interference screening experiments identified numerous NUPs as potential nuclear transport factors for the Retroviridae family, including NUP358/RANBP2, NUP153, NUP98, and NUP214/ CAN (NUP 50,62,85,107,133,155,160,210, ELYS, and TRP) (Ebina et al 2004;Brass et al 2008;König et al 2008;Woodward et al 2009;Lee et al 2010;Zhang et al 2010;Engelman 2011, 2013a;Di Nunzio et al 2013;Matreyek et al 2013b). Among them, NUP358 and NUP214 are located exclusively at the cytoplasmic face.…”

Section: Nucleoporins (Nups)mentioning

confidence: 99%

“…Depletion or overexpression of Tpr causes remodeling of chromatin proximal to the NPC by stabilization of lens epithelium-derived growth factor (LEDGF) and changes the integration site preference (Lelek et al 2015). NUP153 binds directly with HIV-1 IN, CA, and Vpr in an importin ␣-and (or) importin ␤-independent manner for import (Varadarajan et al 2005;Woodward et al 2009;Di Nunzio et al 2013), but CA import may be NUP153 and (or) TNPO3 dependent (Paulillo et al 2005;Cardarelli et al 2012;Matreyek et al 2013b). PF74 and (or) CPSF6 compete with NUP153 for binding to the HIV-1 CA pocket, thus higher concentrations of PF74 can block HIV-1 infection.…”

Section: Nucleoporins (Nups)mentioning

confidence: 99%

“…Most recently, compartmentalization of CA in the nucleus of infected cells was determined by direct interaction with the FG-repeat enriched NUP153 (Matreyek et al 2013b). CA binds with several cellular proteins, including TNPO3, NUP358, NUP153, NUP158, TRIM5␣, CPSF6, and CypA, all of which are involved in nuclear transport machineries (Bieniasz 2012;Matreyek et al 2013b). Because CA does not possess its own NLS, it may provide anchoring sites or pockets for PIC proteins, allowing transport of PIC into the nu-cleus.…”

Section: Gagmentioning

confidence: 99%

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Cellular and viral determinants of retroviral nuclear entry

2016

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Retroviruses must integrate their cDNA into the host genome to generate proviruses. Viral DNA-protein complexes interact with cellular proteins and produce pre-integration complexes, which carry the viral genome and cross the nuclear pore channel to enter the nucleus and integrate viral DNA into host chromosomal DNA. If the reverse transcripts fail to integrate, linear or circular DNA species such as 1-and 2-long terminal repeats are generated. Such complexes encounter numerous cellular proteins in the cytoplasm, which restrict viral infection and protect the nucleus. To overcome host cell defenses, the pathogens have evolved several evasion strategies. Viral proteins often contain nuclear localization signals, allowing entry into the nucleus. Among more than 1000 proteins identified as required for HIV infection by RNA interference screening, karyopherins, cleavage and polyadenylation specific factor 6, and nucleoporins have been predominantly studied. This review discusses current opinions about the synergistic relationship between the viral and cellular factors involved in nuclear import, with focus on the unveiled mysteries of the host-pathogen interaction, and highlights novel approaches to pinpoint therapeutic targets.

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“…Both nucleoporins are required for HIV-1 nuclear entry as their depletion decreases HIV-1 integration into the host genome (Konig et al, 2008;Di Nunzio et al, 2012). Nup153 and Nup358 can interact with the HIV-1 capsid (Schaller et al, 2011;Di Nunzio et al, 2013b;Matreyek et al, 2013) and Nup358 may serve as a docking site for the HIV-1 preintegration complex (PIC) at the nuclear pore, while Nup153 might play a role in the nuclear entry of the PIC (Lee et al, 2010;Matreyek and Engelman, 2011). Nup153 may interact with PICs docked on Nup358 and facilitate their transport through the NPC.…”

Section: Nup153 and Hivmentioning

confidence: 99%

The nuclear pore complex: structure and function

Duhéron¹,

Fahrenkrog²

2017

Atlas of Genetics and Cytogenetics in Oncology and Haematology

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Forkhead box P3 (FOXP3), a gene member of the forkhead/winged-helix family of transcription regulators, is Nuclear pore complexes (NPCs) are large multi-protein complexes, which are embedded in the nuclear envelope and which are regulating the molecular exchange between the nucleus and the cytoplasm. While electron microscopy and cryo-electron tomography studies have provided high-resolution pictures of the NPC structure as entity, the challenge nowadays is to elucidate the organization and the functions of nuclear pore proteins (nucleoporins or Nups) inside and outside the NPC. Nucleoporins are not only involved in nucleocytoplasmic transport, but in an increasing number of other cellular processes, such as kinetochore organization, cell cycle regulation, DNA repair, and gene expression. The implication of nucleoporins in these diverse processes links them also to a wide variety of human diseases, such as cancer and autoimmune diseases. Here we review the progress made in defining the molecular arrangement of nucleoporins within the NPC and use the example of Nup153 to illustrate the versatility of individual nucleoporins and their implication in various human diseases.

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Nucleoporin NUP153 Phenylalanine-Glycine Motifs Engage a Common Binding Pocket within the HIV-1 Capsid Protein to Mediate Lentiviral Infectivity

Cited by 249 publications

References 90 publications

HIV-1 Resistance to the Capsid-Targeting Inhibitor PF74 Results in Altered Dependence on Host Factors Required for Virus Nuclear Entry

HIV-1 Resistance to the Capsid-Targeting Inhibitor PF74 Results in Altered Dependence on Host Factors Required for Virus Nuclear Entry

Cellular and viral determinants of retroviral nuclear entry

The nuclear pore complex: structure and function

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