Human papillomavirus infection requires the TSG101 component of the ESCRT machinery

Broniarczyk, Justyna; Bergant, Martina; Goździcka-Józefiak, Anna; Banks, Lawrence

doi:10.1016/j.virol.2014.05.005

Cited by 32 publications

(42 citation statements)

References 37 publications

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“…The transit of vesicles to the TGN and nuclear entry probably involves movement along microtubules by way of molecular motors, since L2 has been shown to interact with dynein [25], while a membrane-associated cellular protease g-secretase is required for localization of L2 and viral DNA to the Golgi apparatus and endoplasmic reticulum [17,26]. It is becoming clear that extensive cellular cargo sorting machinery is involved in L2-related HPV trafficking from endosomes to the nucleus, including SNX17 [21] and SNX27 [27], retromer proteins [23], and at least one member of the ESCRT complex, TSG101 [28]. The exact sequence of events in which HPVs utilize members of the host sorting machinery for the transition to TGN and nucleus remains an important question.…”

Section: Introductionmentioning

confidence: 99%

Characterizing the spatio-temporal role of sorting nexin 17 in human papillomavirus trafficking

Bergant

Peternel

Pim

et al. 2017

Journal of General Virology

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The human papillomavirus (HPV) L2 capsid protein plays an essential role during the early stages of viral infection. Previous studies have shown that the interaction between HPV L2 and endosomal sorting nexin 17 (SNX17) is conserved across multiple PV types where it plays an essential role in infectious entry, suggesting an evolutionarily conserved pathway of PV trafficking. Here we show that the peak time of interaction between HPV-16 L2 and SNX17 is rather early, at 2 h postinfection. Interestingly, the L2-SNX17 interaction appears to be important for facilitating capsid disassembly and L1 dissociation, suggesting that L2 recruitment of SNX17 occurs prior to capsid disassembly. Furthermore, we also found evidence of L2-SNX17 association at the later stages of infectious entry, suggesting that the SNX17-mediated sorting machinery is either involved at different stages of HPV trafficking or that L2-SNX17 interaction is a long-lasting event in HPV trafficking.

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Section: Introductionmentioning

confidence: 99%

Characterizing the spatio-temporal role of sorting nexin 17 in human papillomavirus trafficking

Bergant

Peternel

Pim

et al. 2017

Journal of General Virology

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“…DiGiuseppe and coworkers revealed that L2 amino acid residues 64 to 81 and 163 to 170 and the L2 C-terminal exposure on the cytosolic side of intracellular membranes enable interaction with cytosolic host cell factors (39). Interactions of L2 with actin (40), components of the retrograde transport machinery (37,41,42), sorting nexins 17 and 27, TSG101, ␥-secretase, and Hsc70, as well as the microtubule network, have been reported (37,(41)(42)(43)(44)(45)(46)(47)(48). These interactions result in trafficking to the Golgi network (37,41,42,47), transport toward the nucleus (43,44), and accumulation at nuclear substructures (49)(50)(51)(52)(53).…”

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confidence: 99%

The Cytoskeletal Adaptor Obscurin-Like 1 Interacts with the Human Papillomavirus 16 (HPV16) Capsid Protein L2 and Is Required for HPV16 Endocytosis

Wüstenhagen

Hampe

Boukhallouk

et al. 2016

J Virol

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The human papillomavirus (HPV) capsid protein L2 is essential for viral entry. To gain a deeper understanding of the role of L2, we searched for novel cellular L2-interacting proteins. A yeast two-hybrid analysis uncovered the actin-depolymerizing factor gelsolin, the membrane glycoprotein dysadherin, the centrosomal protein 68 (Cep68), and the cytoskeletal adaptor protein obscurin-like 1 protein (OBSL1) as putative L2 binding molecules. Pseudovirus (PsV) infection assays identified OBSL1 as a host factor required for gene transduction by three oncogenic human papillomavirus types, HPV16, HPV18, and HPV31. In addition, we detected OBSL1 expression in cervical tissue sections and noted the involvement of OBSL1 during gene transduction of primary keratinocytes by HPV16 PsV. Complex formation of HPV16 L2 with OBSL1 was demonstrated in coimmunofluorescence and coimmunoprecipitation studies after overexpression of L2 or after PsV exposure. We observed a strong colocalization of OBSL1 with HPV16 PsV and tetraspanin CD151 at the plasma membrane, suggesting a role for OBSL1 in viral endocytosis. Indeed, viral entry assays exhibited a reduction of viral endocytosis in OBSL1-depleted cells. Our results suggest OBSL1 as a novel L2-interacting protein and endocytosis factor in HPV infection. IMPORTANCEHuman papillomaviruses infect mucosal and cutaneous epithelia, and the high-risk HPV types account for 5% of cancer cases worldwide. As recently discovered, HPV entry occurs by a clathrin-, caveolin-, and dynamin-independent endocytosis via tetraspanin-enriched microdomains. At present, the cellular proteins involved in the underlying mechanism of this type of endocytosis are under investigation. In this study, the cytoskeletal adaptor OBSL1 was discovered as a previously unrecognized interaction partner of the minor capsid protein L2 and was identified as a proviral host factor required for HPV16 endocytosis into target cells. The findings of this study advance the understanding of a so far less well-characterized endocytic pathway that is used by oncogenic HPV subtypes. Human papillomaviruses (HPVs) are small, nonenveloped DNA viruses that infect dividing basal keratinocytes of skin and mucosa via microlesions of the tissue. HPV is capable of inducing benign epithelial warts on the skin and mucosa, and infection with a high-risk HPV type may cause cervical and other anogenital and oropharyngeal cancers (1, 2). Cervical cancer is the third most common cancer in women worldwide and is associated with HPV infection, more precisely with high-risk HPV types such as HPV16, HPV18, and HPV31 (3). HPV is composed of a viral capsid with the major capsid protein L1, the minor capsid protein L2, and the viral genome. One icosahedral capsid contains 360 copies of L1, which can self-assemble to 72 pentamers, and up to 72 copies of the minor capsid protein L2, located inside the L1 shell (4-6). The capsid proteins L1 and L2 are key players in early events of infection, such as virus binding at the plasma membrane, cell entry, and t...

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“…Previous studies had shown that the ESCRT component TSG101 and ESCRT-associated ALIX were required for infectious entry of HPV-163031. We were interested first in investigating whether the ESCRT machinery was required for infection with other papillomavirus (PV) types and, further, how the ESCRT machinery might play a role in this process.…”

Section: Resultsmentioning

confidence: 99%

“…To do this we made use of the Pseudovirion (PsVs) system, where L1 and L2 are assembled into PsVs in HEK293TT cells and package a luciferase reporter plasmid35. In this study we used HPV-16, HPV-5, BPV-1, MmuPV-1 and SfPV-1, and as a control we also included Merkel Cell Polyomavirus (MCPyV) in the assays, which in our previous assays was found to be unaffected by TSG101 depletion30. HaCaT cells were first transfected with siRNAs against TSG101, VPS4A, VPS4B and HRS and after 72 h the cells were infected with the different PsVs containing luciferase.…”

Section: Resultsmentioning

confidence: 99%

“…Analysis of the location of incoming HPV capsids had suggested their presence within MVBs12, and studies had also shown a potential requirement for the ESCRT component TSG101 for efficient virus entry30. More recently, the ESCRT associated adaptor protein ALIX, was also shown to be required for intracellular trafficking of incoming HPVs and subsequent capsid disassembly31.…”

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confidence: 99%

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The VPS4 component of the ESCRT machinery plays an essential role in HPV infectious entry and capsid disassembly

Broniarczyk

Pim

Massimi

et al. 2017

Sci Rep

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Human Papillomavirus (HPV) infection involves multiple steps, from cell attachment, through endocytic trafficking towards the trans-Golgi network, and, ultimately, the entry into the nucleus during mitosis. An essential viral protein in infectious entry is the minor capsid protein L2, which engages different components of the endocytic sorting machinery during this process. The ESCRT machinery is one such component that seems to play an important role in the early stages of infection. Here we have analysed the role of specific ESCRT components in HPV infection, and we find an essential role for VPS4. Loss of VPS4 blocks infection with multiple PV types, suggesting an evolutionarily conserved critical step in infectious entry. Intriguingly, both L1 and L2 can interact with VPS4, and appear to be in complex with VPS4 during the early stages of virus infection. By using cell lines stably expressing a dominant-negative mutant form of VPS4, we also show that loss of VPS4 ATPase activity results in a marked delay in capsid uncoating, resulting in a defect in the endocytic transport of incoming PsVs. These results demonstrate that the ESCRT machinery, and in particular VPS4, plays a critical role in the early stages of PV infection.

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Human papillomavirus infection requires the TSG101 component of the ESCRT machinery

Cited by 32 publications

References 37 publications

Characterizing the spatio-temporal role of sorting nexin 17 in human papillomavirus trafficking

Characterizing the spatio-temporal role of sorting nexin 17 in human papillomavirus trafficking

The Cytoskeletal Adaptor Obscurin-Like 1 Interacts with the Human Papillomavirus 16 (HPV16) Capsid Protein L2 and Is Required for HPV16 Endocytosis

The VPS4 component of the ESCRT machinery plays an essential role in HPV infectious entry and capsid disassembly

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