scholarly journals γ-Secretase promotes membrane insertion of the human papillomavirus L2 capsid protein during virus infection

2018 ◽  
Vol 217 (10) ◽  
pp. 3545-3559 ◽  
Author(s):  
Takamasa Inoue ◽  
Pengwei Zhang ◽  
Wei Zhang ◽  
Kylia Goodner-Bingham ◽  
Allison Dupzyk ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Retrograde Transport ◽  
Hpv Infection ◽  
Human Papillomaviruses ◽  
Membrane Insertion ◽  
Human Pathogens ◽  
Transport Pathway ◽  
Chaperone Function ◽  
Endosomal Membranes

Despite their importance as human pathogens, entry of human papillomaviruses (HPVs) into cells is poorly understood. The transmembrane protease γ-secretase executes a crucial function during the early stages of HPV infection, but the role of γ-secretase in infection and the identity of its critical substrate are unknown. Here we demonstrate that γ-secretase harbors a previously uncharacterized chaperone function, promoting low pH–dependent insertion of the HPV L2 capsid protein into endosomal membranes. Upon membrane insertion, L2 recruits the cytosolic retromer, which enables the L2 viral genome complex to enter the retrograde transport pathway and traffic to the Golgi en route for infection. Although a small fraction of membrane-inserted L2 is also cleaved by γ-secretase, this proteolytic event appears dispensable for HPV infection. Our findings demonstrate that γ-secretase is endowed with an activity that can promote membrane insertion of L2, thereby targeting the virus to the productive infectious pathway.

scholarly journals Papillomaviruses Go Retro

Pathogens ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 267
Author(s):  
Jian Xie ◽  
Pengwei Zhang ◽  
Mac Crite ◽  
Daniel DiMaio
Keyword(s):  
Retrograde Transport ◽  
Hpv Infection ◽  
Human Papillomaviruses ◽  
Cellular Biology ◽  
Genome Replication ◽  
Intracellular Membrane ◽  
Transport Pathway ◽  
Hpv Dna ◽  
Membrane Bound ◽  
Entry Mechanism

Human papillomaviruses are important pathogens responsible for approximately 5% of cancer as well as other important human diseases, but many aspects of the papillomavirus life cycle are poorly understood. To undergo genome replication, HPV DNA must traffic from the cell surface to the nucleus. Recent findings have revolutionized our understanding of HPV entry, showing that it requires numerous cellular proteins and proceeds via a series of intracellular membrane-bound vesicles that comprise the retrograde transport pathway. This paper reviews the evidence supporting this unique entry mechanism with a focus on the crucial step by which the incoming virus particle is transferred from the endosome into the retrograde pathway. This new understanding provides novel insights into basic cellular biology and suggests novel rational approaches to inhibit HPV infection.

scholarly journals Retromer stabilizes transient membrane insertion of L2 capsid protein during retrograde entry of human papillomavirus

2021 ◽  
Vol 7 (27) ◽  
pp. eabh4276
Author(s):  
Jian Xie ◽  
Pengwei Zhang ◽  
Mac Crite ◽  
Christina V. Lindsay ◽  
Daniel DiMaio
Keyword(s):  
Capsid Protein ◽  
Protein Trafficking ◽  
Transmembrane Proteins ◽  
Cellular Protein ◽  
Human Papillomaviruses ◽  
Genetic Interactions ◽  
Cell Penetrating Peptide ◽  
Membrane Insertion ◽  
Associated Proteins ◽  
A Cell

Retromer, a cellular protein trafficking complex, sorts human papillomaviruses (HPVs) into the retrograde pathway for transport of HPV to the nucleus during virus entry. Here, we conducted a protein modulation screen to isolate four artificial transmembrane proteins called traptamers that inhibit different steps of HPV entry. By analyzing cells expressing pairs of traptamers, we ordered the trafficking steps during entry into a coherent pathway. One traptamer stimulates ubiquitination of the L2 capsid protein or associated proteins and diverts incoming virus to the lysosome, whereas the others act downstream by preventing sequential passage of the virus through retrograde compartments. Complex genetic interactions between traptamers revealed that a cell-penetrating peptide (CPP) on L2 mediates transient insertion of L2 into the endosome membrane, which is stabilized by retromer-L2 binding. These results define the retrograde entry route taken by HPV and show that retromer can play a role in CPP-mediated membrane insertion.

scholarly journals Epigenetic Regulation of the Human Papillomavirus Life Cycle

Pathogens ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 483 ◽  
Author(s):  
Michelle Mac ◽  
Cary A. Moody
Keyword(s):  
Life Cycle ◽  
Hpv Infection ◽  
Epigenetic Modifications ◽  
Human Papillomaviruses ◽  
Epigenetic Changes ◽  
Epigenetic Alterations ◽  
Malignant Phenotype ◽  
Public Health Burden ◽  
Damage Response

Persistent infection with certain types of human papillomaviruses (HPVs), termed high risk, presents a public health burden due to their association with multiple human cancers, including cervical cancer and an increasing number of head and neck cancers. Despite the development of prophylactic vaccines, the incidence of HPV-associated cancers remains high. In addition, no vaccine has yet been licensed for therapeutic use against pre-existing HPV infections and HPV-associated diseases. Although persistent HPV infection is the major risk factor for cancer development, additional genetic and epigenetic alterations are required for progression to the malignant phenotype. Unlike genetic mutations, the reversibility of epigenetic modifications makes epigenetic regulators ideal therapeutic targets for cancer therapy. This review article will highlight the recent advances in the understanding of epigenetic modifications associated with HPV infections, with a particular focus on the role of these epigenetic changes during different stages of the HPV life cycle that are closely associated with activation of DNA damage response pathways.

scholarly journals Broad Neutralization Responses Against Oncogenic Human Papillomaviruses Induced by a Minor Capsid L2 Polytope Genetically Incorporated Into Bacterial Ferritin Nanoparticles

2020 ◽  
Vol 11 ◽  
Author(s):  
Fan Yang ◽  
Filipe C. Mariz ◽  
Xueer Zhao ◽  
Gloria Spagnoli ◽  
Simone Ottonello ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Capsid Protein ◽  
Pyrococcus Furiosus ◽  
Neutralizing Antibody ◽  
Hpv Infection ◽  
Cross Reactivity ◽  
Human Papillomaviruses ◽  
Promising Alternative ◽  
Peptide Epitope ◽  
Hpv Types

Cervical cancer remains a global health burden despite the introduction of highly effective vaccines for the prophylaxis of causative human papillomavirus infection (HPV). Current efforts to eradicate cervical cancer focus on the development of broadly protective, cost-effective approaches. HPV minor capsid protein L2 is being recognized as a promising alternative to the major capsid protein L1 because of its ability to induce responses against a wider range of different HPV types. However, a major limitation of L2 as a source of cross-neutralizing epitopes is its lower immunogenicity compared to L1 when assembled into VLPs. Various approaches have been proposed to overcome this limitation, we developed and tested ferritin-based bio-nanoparticles displaying tandemly repeated L2 epitopes from eight different HPV types grafted onto the surface of Pyrococcus furiosus thioredoxin (Pf Trx). Genetic fusion of the Pf Trx-L2(8x) module to P. furiosus ferritin (Pf Fe) did not interfere with ferritin self-assembly into an octahedral structure composed by 24 protomers. In guinea pigs and mice, the ferritin super-scaffolded, L2 antigen induced a broadly neutralizing antibody response covering 14 oncogenic and two non-oncogenic HPV types. Immune-responsiveness lasted for at least one year and the resulting antibodies also conferred protection in a cervico-vaginal mouse model of HPV infection. Given the broad organism distribution of thioredoxin and ferritin, we also verified the lack of cross-reactivity of the antibodies elicited against the scaffolds with human thioredoxin or ferritin. Altogether, the results of this study point to P. furiosus ferritin nanoparticles as a robust platform for the construction of peptide-epitope-based HPV vaccines.

scholarly journals Cell-penetrating peptide inhibits retromer-mediated human papillomavirus trafficking during virus entry

2020 ◽  
Vol 117 (11) ◽  
pp. 6121-6128 ◽  
Author(s):  
Pengwei Zhang ◽  
Ruben Moreno ◽  
Paul F. Lambert ◽  
Daniel DiMaio
Keyword(s):  
Human Papillomavirus ◽  
Virus Replication ◽  
Protein Interactions ◽  
Retrograde Transport ◽  
Hpv Infection ◽  
Cellular Protein ◽  
Protein Protein Interactions ◽  
Transport Pathway ◽  
Viral Genomes ◽  
Cell Penetrating

Virus replication requires critical interactions between viral proteins and cellular proteins that mediate many aspects of infection, including the transport of viral genomes to the site of replication. In human papillomavirus (HPV) infection, the cellular protein complex known as retromer binds to the L2 capsid protein and sorts incoming virions into the retrograde transport pathway for trafficking to the nucleus. Here, we show that short synthetic peptides containing the HPV16 L2 retromer-binding site and a cell-penetrating sequence enter cells, sequester retromer from the incoming HPV pseudovirus, and inhibit HPV exit from the endosome, resulting in loss of viral components from cells and in a profound, dose-dependent block to infection. The peptide also inhibits cervicovaginal HPV16 pseudovirus infection in a mouse model. These results confirm the retromer-mediated model of retrograde HPV entry and validate intracellular virus trafficking as an antiviral target. More generally, inhibiting virus replication with agents that can enter cells and disrupt essential protein-protein interactions may be applicable in broad outline to many viruses.

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

2016 ◽  
Vol 90 (23) ◽  
pp. 10629-10641 ◽  
Author(s):  
Elena Wüstenhagen ◽  
Laura Hampe ◽  
Fatima Boukhallouk ◽  
Marc A. Schneider ◽  
Gilles A. Spoden ◽  
...  
Keyword(s):  
Human Papillomavirus ◽  
Capsid Protein ◽  
Viral Entry ◽  
Adaptor Protein ◽  
Hpv Infection ◽  
Host Factor ◽  
Human Papillomaviruses ◽  
Endocytic Pathway ◽  
Target Cells ◽  
Gene Transduction

ABSTRACTThe 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.

scholarly journals Episome partitioning and symmetric cell divisions: Quantifying the role of random events in the persistence of HPV infections

2021 ◽  
Vol 17 (9) ◽  
pp. e1009352
Author(s):  
Thomas Beneteau ◽  
Christian Selinger ◽  
Mircea T. Sofonea ◽  
Samuel Alizon
Keyword(s):  
Hpv Infection ◽  
Human Papillomaviruses ◽  
Relative Importance ◽  
Oncogenic Viruses ◽  
Sexually Transmitted ◽  
Cell Divisions ◽  
Random Events ◽  
Stem Cell Division ◽  
Underlying Mechanisms

Human Papillomaviruses (HPV) are one of the most prevalent sexually transmitted infections (STI) and the most oncogenic viruses known to humans. The vast majority of HPV infections clear in less than 3 years, but the underlying mechanisms, especially the involvement of the immune response, are still poorly known. Building on earlier work stressing the importance of randomness in the type of cell divisions in the clearance of HPV infection, we develop a stochastic mathematical model of HPV dynamics that combines the previous aspect with an explicit description of the intracellular level. We show that the random partitioning of virus episomes upon stem cell division and the occurrence of symmetric divisions dramatically affect viral persistence. These results call for more detailed within-host studies to better understand the relative importance of stochasticity and immunity in HPV infection clearance.

scholarly journals Immunohistochemical Detection of early and late Human Papillomaviruses (HPV) proteins in Retinoblastoma tumor

2020 ◽  
Author(s):  
Nara Diniz Soares Pessoa ◽  
Thatiana Correia Melo ◽  
Rodrigo Pinheiro Araldi ◽  
Rofrigo Franco Carvalho ◽  
Willy Becak ◽  
...  
Keyword(s):  
Hpv Infection ◽  
Human Papillomaviruses ◽  
Immunohistochemical Detection ◽  
Hpv Dna ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Retinoblastoma Tumor ◽  
Formalin Fixed

In this study, we evaluated the presence of early and late Human Papillomavirus (HPV) proteins in retinoblastoma Brazilian patients. For this, 8 formalin-fixed paraffin-embedded retinoblastoma tissue blocks were used. HPV DNA presence was determined by in situ hybridization (ISH). Immunohistochemistry were performed to verify HPV16/18 E6, E1^E4, and L1 proteins. HPV was detected in all retinoblastoma tumors and viral DNA was labeled in tumor cells, retinal layers and optical nerve structures. In addition, E1^E4, E6 and L1 proteins were detected in all samples in the same areas where HPV DNA was detected. Our data showed the presence and expression of early and late HPV proteins in retinoblastoma tumors from Brazilian children. However, further studies should be performed to clarify the role of HPV infection in retinoblastoma tumor.

scholarly journals Detection of Human Papillomaviruses (HPVs) by Immunohistochemistry (P16ink4a) In Verrucous Skin Lesions

2021 ◽  
pp. 57-68
Author(s):  
Shone Thomas Babu ◽  
Mary Lilly
Keyword(s):  
Risk Factors ◽  
Risk Factor ◽  
Skin Lesions ◽  
Hpv Infection ◽  
Squamous Cell Carcinomas ◽  
Human Papillomaviruses ◽  
Cervical Lesions ◽  
Malignant Lesions ◽  
P16 Expression

Even though the p16 marker allows us to distinguish between benign and malignant verrucous skin lesions, we are unable to determine the patient's Human Papillomaviruses (HPVs) status despite the existence of recognized histopathological features that indicate its presence like koilocytic change, since a proper correlation couldn’t be made between this feature and p16 expression. Even though PV type 16 and 18 are a risk factor for developments of anogenital skin lesion diffuse p16 expression cannot always be attributed to HPVs as there may be several other risk factors causing skin lesions, unlike in cervical lesions such as squamous cell carcinomas, many studies have established the role of oncogenic HPVs with its carcinogenesis. This marker cannot be used as a surrogate for detection of HPV infection. The present study was the expression of p16INK4A in histological sections of verrucous skin lesions. To compare the expression of p16INK4a in benign, premalignant and malignant lesions involving the  skin and  comparing the  pattern of expression of p16INK4a in skin lesions  by  immunohistochemistry  and correlating the results with certain histological parameters that might indicate HPV infection.

scholarly journals Cytokine response following perturbation of the cervicovaginal milieu during HPV genital infection

2021 ◽  
Author(s):  
Christian Selinger ◽  
Massilva Rahmoun ◽  
Carmen Lia Murall ◽  
Claire Bernat ◽  
Vanina Boué ◽  
...  
Keyword(s):  
Hpv Infection ◽  
Human Papillomaviruses ◽  
Sexually Active ◽  
Oncogenic Viruses ◽  
Cytokine Network ◽  
Cervical Cancers ◽  
Effector Molecules ◽  
Inflammatory Proteins ◽  
Factor Modeling

AbstractHuman papillomaviruses (HPVs) are the most oncogenic viruses known to human, causing nearly all cervical cancers worldwide. Highly prevalent in young, sexually active women, most HPV infections are cleared within 3 years, and only a minority of those infections persist and lead to cancer later in life. To better characterize the immuno-modulatory impact of early HPV infection and more generally perturbations of the cervicovaginal milieu, we measured changes in a panel of 20 cytokines, known as highly dynamic effector molecules implicated in cell signaling. We analyzed 92 cervicovaginal samples collected from young, sexually active women who were tested for or diagnosed with HPV, chlamydia, and bacterial vaginosis. Also, symptoms associated with genital inflammation and infection were collected through self-reporting.Following a parsimonious multi-factor modeling approach, our statistical analyses revealed that increased IL-1Alpha and IL-12/IL-23p40 concentrations were associated with HPV infection. Cytokine network analysis further highlighted the role of IL-1Alpha and macrophage inflammatory proteins (MIP-3Alpha) in HPV-associated immuno-modulation.

Sign in / Sign up

Export Citation Format

Share Document