Polyethylenimine Is a Strong Inhibitor of Human Papillomavirus and Cytomegalovirus Infection

Spoden, Gilles A.; Besold, Katrin; Krauter, Steffi; Plachter, Bodo; Hanik, Nils; Kilbinger, Andreas F. M.; Lambert, Carsten; Florin, Luise

doi:10.1128/aac.05147-11

Cited by 57 publications

(64 citation statements)

References 56 publications

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“…In contrast to reports suggesting OBSL1 localization at the Golgi apparatus (65), no colocalization between the Golgi and OBSL1 was detected in HaCaT cells. Instead, we observed association of OBSL1 with the actin cytoskeleton, the tetraspanin CD151-both required for papillomavirus endocytosis-and colocalization of OBSL1 and L2 at the plasma membrane during HPV16 PsV entry, suggesting a role for OBSL1 in early events of HPV16 entry.…”

Section: Discussioncontrasting

confidence: 52%

“…Quantitative Western blot analysis demonstrated that siRNA-mediated OBSL1 depletion had no or a minor effect on initial HPV16 cell surface binding. As an internal control, HaCaT cells were transfected with control siRNA and treated with polyethyleneimine (PEI), a strong inhibitor for primary HPV16 PsV binding (65). These data were strengthened by results obtained from L1 surface staining by flow cytometry analysis (Fig.…”

Section: Hpv16 L2 Forms a Complex With Obsl1mentioning

confidence: 57%

“…HaCaT cells were grown in 24-well plates and transfected with target-specific siRNA for 48 h. Afterwards, cells were detached with 0.05% trypsin-2.5 mM EDTA, resuspended in DMEM, and transferred into siliconized reaction tubes. Control cells were pretreated with polyethyleneimine (PEI), an inhibitor of HPV infection (SigmaAldrich, St. Louis, MO), for 1 h at 4°C as indicated below and described previously (65). Subsequently, all cells were exposed to 200 to 500 HPV16 PsVs for 1 h at 4°C on an overhead rotator.…”

Section: Methodsmentioning

confidence: 99%

“…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%

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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: Discussioncontrasting

confidence: 52%

Section: Hpv16 L2 Forms a Complex With Obsl1mentioning

confidence: 57%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

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 expression of the reporter is used as the readout for successful cell entry after delivery of the viral pseudogenome to the nucleus. Luciferase was used as the reporter in this study, and PsVs were prepared as described previously (14,15).…”

mentioning

confidence: 99%

Human Papillomavirus Types 16, 18, and 31 Share Similar Endocytic Requirements for Entry

Spoden

Kühling

Cordes

et al. 2013

J Virol

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Human papillomavirus type 18 (HPV18), one of the HPVs with malignant potential, enters cells by an unknown endocytic mechanism. The key cellular requirements for HPV18 endocytosis were tested in comparison to those for HPV16 and -31 endocytoses. HPV18 (like HPV16 and -31) entry was independent of clathrin, caveolin, dynamin, and lipid rafts but required actin polymerization and tetraspanin CD151, and the viruses were routed to the same LAMP-1-positive compartment. Hence, the viruses shared similar cellular requirements for endocytic entry. The human papillomaviruses (HPVs) are a family of nonenveloped DNA viruses with transforming potential. A particular subgroup of HPVs, the high-risk types, are additionally associated with cervical and anogenital cancers. Of those high-risk types, HPV type 16 (HPV16) is the most prevalent and the best studied. HPVs enter the basal cells of squamous epithelia by endocytosis during initial infection (1). For cell entry, most viruses employ one of several existing pathways for endocytosis (2, 3). These pathways are typically differentiated by the cellular proteins mediating endocytic vacuole formation, by the ultrastructural morphology of endocytic pits, and by the timing of vesicle formation (2-4). Although HPV16 cell entry was originally attributed to clathrinmediated endocytosis (CME) in various cell types (5-8), it has recently been shown that HPV16 enters cells by a novel ligandinduced endocytic pathway that is clathrin, caveolin, lipid raft, and dynamin independent but that depends on highly regulated actin dynamics and an association with CD151-containing tetraspanin-enriched microdomains (9-11) The high-risk-type HPV31 may differ in the mode of endocytosis from HPV16, as it has been described to enter into human keratinocytes by a caveolin/lipid raft-mediated mechanism (12). However, in COS-7 cells and 293TT cells, HPV16 and -31 endocytoses appear to occur by a similar mechanism that is independent of lipid rafts (5).One of the more-prevalent high-risk types, which has not been analyzed for its mode of endocytic entry, is HPV18. Here, we aimed to delineate the general endocytic requirements for infectious HPV18 endocytosis. In particular, we aimed to analyze whether HPV18 exhibits requirements similar to those of HPV16 or -31 for endocytic entry into host cells. Therefore, we compared the efficacy of cell entry after cell perturbation for the main determinants of several endocytic pathways in HeLa and HaCaT cells. The cell biological determinants that allow differentiation of the different endocytic pathways include clathrin for CME, caveolin for caveolin-mediated mechanisms, cholesterol depletion for lipid raft-dependent mechanisms (e.g., caveolar/lipid raft endocytosis or glycosphingolipid-enriched endocytic carriers), dynamin-2 for dynamin-mediated endocytic processes (e.g., CME, caveolar endocytosis, and the interleukin-2 pathway), and actin polymerization dynamics for actin-mediated endocytic processes (macropinocytosis, caveolar endocytosis, and HPV16 endocytosis) (...

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Broad‐Spectrum Antiviral Peptides and Polymers

Kuroki

Tay

Lee

et al. 2021

Adv Healthcare Materials

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As the human cost of the pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is still being witnessed worldwide, the development of broad-spectrum antiviral agents against emerging and re-emerging viruses is seen as a necessity to hamper the spread of infections. Various targets during the viral life-cycle can be considered to inhibit viral infection, from viral attachment to viral fusion or replication. Macromolecules represent a particularly attractive class of therapeutics due to their multivalency and versatility.Although several antiviral macromolecules hold great promise in clinical applications, the emergence of resistance after prolonged exposure urges the need for improved solutions. In the present article, the recent advancement in the discovery of antiviral peptides and polymers with diverse structural features and antiviral mechanisms is reviewed. Future perspectives, such as, the development of virucidal peptides/polymers and their coatings against SARS-CoV-2 infection, standardization of antiviral testing protocols, and use of artificial intelligence or machine learning as a tool to accelerate the discovery of antiviral macromolecules, are discussed.

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Polyethylenimine Is a Strong Inhibitor of Human Papillomavirus and Cytomegalovirus Infection

Cited by 57 publications

References 56 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

Human Papillomavirus Types 16, 18, and 31 Share Similar Endocytic Requirements for Entry

Broad‐Spectrum Antiviral Peptides and Polymers

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