An L2 SUMO interacting motif is important for PML localization and infection of human papillomavirus type 16

Bund, Timo; Spoden, Gilles A.; Koynov, Kaloian; Hellmann, Nadja; Boukhallouk, Fatima; Arnold, Philipp; Hinderberger, Dariush; Florin, Luise

doi:10.1111/cmi.12271

Cited by 29 publications

(60 citation statements)

References 94 publications

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“…HPV16, -18, and -31 pseudoviruses (PsVs) and HPV16 L1-only PsV were prepared as previously described and published (33,53,64). Quantification of the reporter gene plasmid was performed as published before (28,53,65).…”

Section: Methodsmentioning

confidence: 99%

“…Fixed cells were washed three times with PBS and blocked for 30 min with PBS-1% (wt/vol) bovine serum albumin (BSA; AppliChem, Darmstadt, Germany). For L2 staining after PsV exposure, cells were incubated with Click-iT reaction cocktail according to the manufacturer's instructions (without the addition of EdU label for PsV staining) as described previously (22,53). Afterwards, proteins were stained with the desired antibodies at 37°C for 1 h. After washing with PBS, coverslips were blocked with PBS-1% BSA for 10 min and subsequently incubated with Alexa-conjugated specific secondary antibodies or phalloidin-tetramethyl rhodamine isothiocyanate (TRITC; Sigma-Aldrich, St. Louis, MO) for 1 h at 37°C in the dark.…”

Section: Methodsmentioning

confidence: 99%

“…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). Furthermore, L2 possesses a membrane-destabilizing peptide in its C terminus (54) and a transmembrane domain in its N terminus (55) that are both important for translocation to the cytoplasm.…”

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confidence: 99%

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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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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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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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“…Several viruses relocate viral proteins into ND10 bodies to enhance their replication. Thus, SV-40 large T antigen colocalizes with PML and ND10 bodies, and papillomavirus minor capsid protein L2 colocalizes with PML and ND10 (24)(25)(26). It has been reported that one component of ND10, SP100, provides intrinsic immunity against papilloma viruses (27).…”

mentioning

confidence: 98%

The SP100 component of ND10 enhances accumulation of PML and suppresses replication and the assembly of HSV replication compartments

Roizman

2017

Proc. Natl. Acad. Sci. U.S.A.

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Nuclear domain 10 (ND10) bodies are small (0.1–1 μM) nuclear structures containing both constant [e.g., promyelocytic leukemia protein (PML), SP100, death domain-associated protein (Daxx)] and variable proteins, depending on the function of the cells or the stress to which they are exposed. In herpes simplex virus (HSV)-infected cells, ND10 bodies assemble at the sites of DNA entering the nucleus after infection. In sequence, the ND10 bodies become viral replication compartments, and ICP0, a viral E3 ligase, degrades both PML and SP100. The amounts of PML and SP100 and the number of ND10 structures increase in cells exposed to IFN-β. Earlier studies have shown that PML has three key functions. Thus, (i) the interaction of PML with viral components facilitates the initiation of replication compartments, (ii) viral replication is significantly less affected by IFN-β in PML−/− cells than in parental PML+/+ cells, and (iii) viral yields are significantly lower in PML−/− cells exposed to low ratios of virus per cell compared with parental PML+/+ cells. This report focuses on the function of SP100. In contrast to PML−/− cells, SP100−/− cells retain the sensitivity of parental SP100+/+ cells to IFN-β and support replication of the ΔICP0 virus. At low multiplicities of infection, wild-type virus yields are higher in SP100−/− cells than in parental HEp-2 cells. In addition, the number of viral replication compartments is significantly higher in SP100−/− cells than in parental SP100+/+ cells or in PML−/− cells.

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“…Using this approach, Greber and co-workers succeeded to produce progeny Ad virions containing labeled genomes and to visualize single genomes of incoming viruses in early phases of infection [29]. This technique has also been used for visualizing incoming genomes of other viruses such as HSV-1 and papillomavirus [30,31,32]. To produce genome-labeled viruses, one may need to test several analogs, since labeling efficiency as well as cellular/viral toxicity varies among them.…”

Section: Experimental Strategies To Study Nuclear Antiviral Factorsmentioning

confidence: 99%

The Role of Nuclear Antiviral Factors against Invading DNA Viruses: The Immediate Fate of Incoming Viral Genomes

Komatsu

Nagata

Wodrich

2016

Viruses

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In recent years, it has been suggested that host cells exert intrinsic mechanisms to control nuclear replicating DNA viruses. This cellular response involves nuclear antiviral factors targeting incoming viral genomes. Herpes simplex virus-1 (HSV-1) is the best-studied model in this context, and it was shown that upon nuclear entry HSV-1 genomes are immediately targeted by components of promyelocytic leukemia nuclear bodies (PML-NBs) and the nuclear DNA sensor IFI16 (interferon gamma inducible protein 16). Based on HSV-1 studies, together with limited examples in other viral systems, these phenomena are widely believed to be a common cellular response to incoming viral genomes, although formal evidence for each virus is lacking. Indeed, recent studies suggest that the case may be different for adenovirus infection. Here we summarize the existing experimental evidence for the roles of nuclear antiviral factors against incoming viral genomes to better understand cellular responses on a virus-by-virus basis. We emphasize that cells seem to respond differently to different incoming viral genomes and discuss possible arguments for and against a unifying cellular mechanism targeting the incoming genomes of different virus families.

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An L2 SUMO interacting motif is important for PML localization and infection of human papillomavirus type 16

Cited by 29 publications

References 94 publications

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

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

The SP100 component of ND10 enhances accumulation of PML and suppresses replication and the assembly of HSV replication compartments

The Role of Nuclear Antiviral Factors against Invading DNA Viruses: The Immediate Fate of Incoming Viral Genomes

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