scholarly journals PDZ Domains and Viral Infection: Versatile Potentials of HPV-PDZ Interactions in relation to Malignancy

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Kazunori Nagasaka ◽  
Kei Kawana ◽  
Yutaka Osuga ◽  
Tomoyuki Fujii
Keyword(s):  
Posttranslational Modifications ◽  
Pdz Domain ◽  
Distribution Patterns ◽  
Spatiotemporal Analysis ◽  
Human Papillomaviruses ◽  
Screening Tools ◽  
Binding Motif ◽  
Normal Epithelium ◽  
Pdz Proteins ◽  
Hpv E6

Cervical cancer is caused by high-risk human papillomaviruses (HPVs), and a unique characteristic of these is a PDZ (P̲SD-95/D̲lg/Z̲O-1-)binding motif in their E6 proteins. Through this motif HPV E6 interacts with a variety of PDZ domain-containing proteins and targets them mainly for degradation. These E6-PDZ interactions exhibit extraordinarily different functions in relation to HPV-induced malignancy, depending upon various cellular contexts; for example, Dlg and Scrib show different distribution patterns from what is seen in normal epithelium, both in localization and in amount, and their loss may be a late-stage marker in malignant progression. Recent studies show that interactions with specific forms of the proteins may have oncogenic potential. In addition, it is interesting that PDZ proteins make a contribution to the stabilization of E6 and viral episomal maintenance during the course of HPV life cycle. Various posttranslational modifications also greatly affect their functions. Phosphorylation of hDlg and hScrib by certain kinases regulates several important signaling cascades, and E6-PDZ interactions themselves are regulated through PKA-dependent phosphorylation. Thus these interactions naturally have great potential for both predictive and therapeutic applications, and, with development of screening tools for identifying novel targets of their interactions, comprehensive spatiotemporal analysis is currently underway.

scholarly journals Recognition of high-risk HPV E6 oncoproteins by 14-3-3 proteins studied by interactomics and crystallography

2020 ◽  
Author(s):  
Gergo Gogl ◽  
Kristina V. Tugaeva ◽  
Pascal Eberling ◽  
Camille Kostmann ◽  
Gilles Trave ◽  
...  
Keyword(s):  
High Risk ◽  
Fluorescence Polarization ◽  
Pdz Domain ◽  
Human Papillomaviruses ◽  
Binding Motif ◽  
Hpv E6 ◽  
Sequence Variations ◽  
Hpv18 E6 ◽  
Isoform Specificity ◽  
High Risk Hpv

AbstractIn tumors induced by high-risk mucosal human papillomaviruses (hrm-HPVs), HPV E6 oncoproteins inhibit apoptotic processes and sustain cell proliferation. E6 from all hrm-HPVs harbor a C-terminal short PDZ domain-binding motif (PBM), whose phosphorylation down-regulates PDZ binding but triggers E6 binding to 14-3-3 proteins. Here we classify PBMs of E6 proteins depending on their principle ability to be phosphorylated and subsequently acquire a 14-3-3-binding motif III consensus, (pS/pT)XX-COOH. Systematic competitive fluorescence polarization measurements show that the PBMs from four selected E6 oncoproteins bind all seven human 14-3-3 isoforms with distinct, wide-ranging affinities, obeying remarkable trends assigned to 14-3-3 isoform specificity and small E6 sequence variations. We crystallized the hrm-HPV18 E6 PBM bound to 14-3-3ζ, revealing a 14-3-3-motif III complex at 1.9 Å resolution. Using fluorescence polarization and crystallography, we also demonstrate that fusicoccin, a molecule that reinforces many known 14-3-3 complexes, destabilizes the 14-3-3-E6 interaction, indicating the druggability of that complex.

scholarly journals Recognition of high-risk HPV E6 oncoproteins by 14-3-3 proteins studied by interactomics and crystallography

2020 ◽  
Author(s):  
Gergő Gógl ◽  
Kristina Tugaeva ◽  
Pascal Eberling ◽  
Camille Kostmann ◽  
Gilles Trave ◽  
...  
Keyword(s):  
High Risk ◽  
Fluorescence Polarization ◽  
Pdz Domain ◽  
Human Papillomaviruses ◽  
Binding Motif ◽  
Hpv E6 ◽  
Sequence Variations ◽  
Hpv18 E6 ◽  
Isoform Specificity ◽  
High Risk Hpv

Abstract In tumors induced by high-risk mucosal human papillomaviruses (hrm-HPVs), HPV E6 oncoproteins inhibit apoptotic processes and sustain cell proliferation. E6 from all hrm-HPVs harbor a C-terminal short PDZ domain-binding motif (PBM), whose phosphorylation down-regulates PDZ binding but triggers E6 binding to 14-3-3 proteins. Here we classify PBMs of E6 proteins depending on their principle ability to be phosphorylated and subsequently acquire a 14-3-3-binding motif III consensus, (pS/pT)XX-COOH. Systematic competitive fluorescence polarization measurements show that the PBMs from four selected E6 oncoproteins bind all seven human 14-3-3 isoforms with distinct, wide-ranging affinities, obeying remarkable trends assigned to 14-3-3 isoform specificity and small E6 sequence variations. We crystallized the hrm-HPV18 E6 PBM bound to 14-3-3ζ, revealing a 14-3-3-motif III complex at 1.9 Å resolution. Using fluorescence polarization and crystallography, we also demonstrate that fusicoccin, a molecule that reinforces many known 14-3-3 complexes, destabilizes the 14-3-3-E6 interaction, indicating the druggability of that complex.

scholarly journals Role of the PDZ Domain-Binding Motif of the Oncoprotein E6 in the Pathogenesis of Human Papillomavirus Type 31

2004 ◽  
Vol 78 (22) ◽  
pp. 12366-12377 ◽  
Author(s):  
Choongho Lee ◽  
Laimonis A. Laimins
Keyword(s):  
High Risk ◽  
Pdz Domain ◽  
Immunohistochemical Analysis ◽  
Human Papillomaviruses ◽  
Binding Motif ◽  
Wild Type ◽  
Pdz Proteins ◽  
Viral Copy Number ◽  
Viral Copy

ABSTRACT A number of PDZ domain-containing proteins have been identified as binding partners for the oncoprotein E6 of the high-risk type human papillomaviruses (HPVs). These include hDlg, hScrib, MAGI-1, MAGI-2, MAGI-3, and MUPP1. The PDZ domain-binding motif (-X-T-X-V) at the carboxy terminus of E6 is essential for targeting PDZ proteins for proteasomal degradation. The presence of this motif only in the high-risk HPVs suggests its possible role in HPV-induced oncogenesis. To investigate the role of the PDZ domain-binding motif of E6 in the HPV life cycle, two mutant HPV31 genomes were constructed: E6ValΔ, with a deletion of the last amino acid residue of E6 (valine), and E6ETQVΔ, with a deletion of the entire PDZ domain-binding motif of E6 (ETQV). Three human foreskin keratinocyte (HFK) cell lines were established which maintained transfected wild-type HPV31 or either of two mutant genomes. Cells containing either of two mutant genomes were significantly retarded in their growth rates and reduced in their viral copy numbers compared to those transfected with wild-type genomes. Western analysis did not reveal any significant changes in the levels of PDZ proteins following stable transfection of any HPV31 genomes into HFKs. Although the E6ETQVΔ-transfected HFKs exhibited a pattern of morphological differentiation that appeared different from the HPV31 wild-type-transfected HFKs in organotypic raft cultures, immunohistochemical analysis failed to identify substantial changes in the differentiation-dependent membrane localization of hDlg proteins. These results suggest that binding of E6 to PDZ proteins modulates the early viral functions such as proliferation and maintenance of the viral copy number in undifferentiated cells.

scholarly journals Multi-PDZ Domain Protein MUPP1 Is a Cellular Target for both Adenovirus E4-ORF1 and High-Risk Papillomavirus Type 18 E6 Oncoproteins

2000 ◽  
Vol 74 (20) ◽  
pp. 9680-9693 ◽  
Author(s):  
Siu Sylvia Lee ◽  
Britt Glaunsinger ◽  
Fiamma Mantovani ◽  
Lawrence Banks ◽  
Ronald T. Javier
Keyword(s):  
High Risk ◽  
Cellular Proliferation ◽  
Pdz Domain ◽  
Viral Proteins ◽  
Cellular Protein ◽  
Human Papillomaviruses ◽  
Binding Motif ◽  
Cellular Factor ◽  
Domain Protein ◽  
Hpv 18

ABSTRACT A general theme that has emerged from studies of DNA tumor viruses is that otherwise unrelated oncoproteins encoded by these viruses often target the same important cellular factors. Major oncogenic determinants for human adenovirus type 9 (Ad9) and high-risk human papillomaviruses (HPV) are the E4-ORF1 and E6 oncoproteins, respectively, and although otherwise unrelated, both of these viral proteins possess a functional PDZ domain-binding motif that is essential for their transforming activity and for binding to the PDZ domain-containing and putative tumor suppressor protein DLG. We report here that the PDZ domain-binding motifs of Ad9 E4-ORF1 and high-risk HPV-18 E6 also mediate binding to the widely expressed cellular factor MUPP1, a large multi-PDZ domain protein predicted to function as an adapter in signal transduction. With regard to the consequences of these interactions in cells, we showed that Ad9 E4-ORF1 aberrantly sequesters MUPP1 within the cytoplasm of cells whereas HPV-18 E6 targets this cellular protein for degradation. These effects were specific because mutant viral proteins unable to bind MUPP1 lack these activities. From these results, we propose that the multi-PDZ domain protein MUPP1 is involved in negatively regulating cellular proliferation and that the transforming activities of two different viral oncoproteins depend, in part, on their ability to inactivate this cellular factor.

scholarly journals Acquisition of a phospho-acceptor site enhances HPV E6 PDZ-binding motif functional promiscuity

2020 ◽  
Vol 101 (9) ◽  
pp. 954-962 ◽  
Author(s):  
Vanessa Sarabia-Vega ◽  
Lawrence Banks
Keyword(s):  
Protein Recognition ◽  
Binding Motif ◽  
Acceptor Site ◽  
Oncogenic Potential ◽  
Pdz Proteins ◽  
Hpv E6 ◽  
Hpv Types ◽  
Pdz Protein ◽  
Minor Sequence ◽  
Hpv 18

All cancer-causing human papillomavirus (HPV) E6 oncoproteins have a C-terminal PDZ-binding motif (PBM), which correlates with oncogenic potential. Nonetheless, several HPVs with little or no oncogenic potential also have an E6 PBM, with minor sequence differences affecting PDZ protein selectivity. Furthermore, certain HPV types have a phospho-acceptor site embedded within the PBM. We therefore compared HPV-18, HPV-66 and HPV-40 E6 proteins to examine the possible link between the ability to target multiple PDZ proteins and the acquisition of a phospho-acceptor site. The mutation of essential residues in HPV-18E6 reduces its phosphorylation, and fewer PDZ substrates are bound. In contrast, the generation of consensus phospho-acceptor sites in HPV-66 and HPV-40 E6 PBMs increases the PDZ proteins recognized. Thus, although phosphorylation of the E6 PBM and PDZ protein recognition are mutually exclusive, they are closely linked, with the acquisition of a phospho-acceptor site also contributing to an expansion in the number of PDZ proteins bound.

scholarly journals The Human Papillomavirus (HPV) E6* Proteins from High-Risk, Mucosal HPVs Can Direct Degradation of Cellular Proteins in the Absence of Full-Length E6 Protein

2009 ◽  
Vol 83 (19) ◽  
pp. 9863-9874 ◽  
Author(s):  
David Pim ◽  
Vjekoslav Tomaić ◽  
Lawrence Banks
Keyword(s):  
High Risk ◽  
Tumor Suppressor ◽  
Pdz Domain ◽  
Direct Interaction ◽  
Human Papillomaviruses ◽  
Full Length ◽  
Cellular Proteins ◽  
Hpv E6 ◽  
E6 Protein ◽  
Hpv 18

ABSTRACT The E6 oncoproteins from high-risk mucosotrophic human papillomaviruses (HPVs) target a range of cellular proteins for proteasome-mediated degradation. Apart from the tumor suppressor p53 and proapoptotic Bcl-2 family member Bak, many targets contain class 1 PDZ domains and are involved in cell junction stability and signaling. The targeting mechanism is considered to function by the E6 protein acting as an adaptor molecule linking a cellular ubiquitin ligase to the target protein. In each case, whether the target is the p53 tumor suppressor or a member of the group of PDZ domain-containing targets, this mechanism relies on a direct interaction between E6 and its cellular target. This study focuses on the impact of the HPV type 18 (HPV-18) E6*I protein on the stability of Akt, Dlg, MAGI-1, MAGI-2, and Scribble. We show that HPV-18 E6* expression can downregulate the expression levels of Akt, Dlg, and Scribble in the absence of full-length HPV-18 E6 protein. The reduction in Dlg levels by E6* is independent of transcription and does not require a direct interaction between the two proteins although the proteasome pathway is involved. Further, we provide evidence that activation of certain signal transduction pathways has a profound effect on the targeting of Dlg by E6* and suggest that high-risk HPV E6 oncoproteins can target certain substrates both directly and indirectly through the E6* proteins and may cooperate in their degradation.

scholarly journals TIP-1 Has PDZ Scaffold Antagonist Activity

2006 ◽  
Vol 17 (10) ◽  
pp. 4200-4211 ◽  
Author(s):  
Christine Alewine ◽  
Olav Olsen ◽  
James B. Wade ◽  
Paul A. Welling
Keyword(s):  
Basolateral Membrane ◽  
Pdz Domain ◽  
Negative Regulator ◽  
Binding Motif ◽  
Type I ◽  
Membrane Expression ◽  
Pdz Proteins ◽  
Protein Protein Interaction ◽  
Multiple Protein ◽  
Interaction Domains

PDZ proteins usually contain multiple protein–protein interaction domains and act as molecular scaffolds that are important for the generation and maintenance of cell polarity and cell signaling. Here, we identify and characterize TIP-1 as an atypical PDZ protein that is composed almost entirely of a single PDZ domain and functions as a negative regulator of PDZ-based scaffolding. We found that TIP-1 competes with the basolateral membrane mLin-7/CASK complex for interaction with the potassium channel Kir 2.3 in model renal epithelia. Consequently, polarized plasma membrane expression of Kir 2.3 is disrupted resulting in pronounced endosomal targeting of the channel, similar to the phenotype observed for mutant Kir 2.3 channels lacking the PDZ-binding motif. TIP-1 is ubiquitously expressed, raising the possibility that TIP-1 may play a similar role in regulating the expression of other membrane proteins containing a type I PDZ ligand.

scholarly journals Protein tyrosine phosphatase H1 is a target of the E6 oncoprotein of high-risk genital human papillomaviruses

2007 ◽  
Vol 88 (11) ◽  
pp. 2956-2965 ◽  
Author(s):  
Stephanie Töpffer ◽  
Andreas Müller-Schiffmann ◽  
Konstantin Matentzoglu ◽  
Martin Scheffner ◽  
Gertrud Steger
Keyword(s):  
High Risk ◽  
Cervical Carcinoma ◽  
Protein Tyrosine Phosphatase ◽  
Carcinoma Cell ◽  
Tyrosine Phosphatase ◽  
Pdz Domain ◽  
Human Papillomaviruses ◽  
Binding Motif ◽  
Protein Tyrosine ◽  
Pdz Domain Proteins

The E6 proteins of high-risk genital human papillomaviruses (HPV), such as HPV types 16 and 18, possess a conserved C-terminal PDZ-binding motif, which mediates interaction with some cellular PDZ domain proteins. The binding of E6 usually results in their ubiquitin-mediated degradation. The ability of E6 to bind to PDZ domain proteins correlates with the oncogenic potential. Using a yeast two-hybrid system, GST pull-down experiments and coimmunoprecipitations, we identified the protein tyrosine phosphatase H1 (PTPH1/PTPN3) as a novel target of the PDZ-binding motif of E6 of HPV16 and 18. PTPH1 has been suggested to function as tumour suppressor protein, since mutational analysis revealed somatic mutations in PTPH1 in a minor fraction of various human tumours. We show here that HPV16 E6 accelerated the proteasome-mediated degradation of PTPH1, which required the binding of E6 to the cellular ubiquitin ligase E6-AP and to PTPH1. The endogenous levels of PTPH1 were particularly low in HPV-positive cervical carcinoma cell lines. The reintroduction of the E2 protein into the HPV16-positive cervical carcinoma cell line SiHa, known to lead to a sharp repression of E6 expression and to induce growth suppression, resulted in an increase of the amount of PTPH1. Our data suggest that reducing the level of PTPH1 may contribute to the oncogenic activity of high-risk genital E6 proteins.

scholarly journals The PDZ Ligand Domain of the Human Papillomavirus Type 16 E6 Protein Is Required for E6's Induction of Epithelial Hyperplasia In Vivo

2003 ◽  
Vol 77 (12) ◽  
pp. 6957-6964 ◽  
Author(s):  
Marie L. Nguyen ◽  
Minh M. Nguyen ◽  
Denis Lee ◽  
Anne E. Griep ◽  
Paul F. Lambert
Keyword(s):  
Amino Acids ◽  
Rat Kidney ◽  
Pdz Domain ◽  
Human Papillomaviruses ◽  
Epithelial Hyperplasia ◽  
Common Amino Acid ◽  
Pdz Proteins ◽  
Binding Domains ◽  
E6 Protein

ABSTRACT Human papillomaviruses (HPVs) are the causative agent of warts. Infections with high-risk HPVs are associated with anogenital and head and neck cancers. One of the viral genes responsible for HPV's oncogenic activity is E6. Mice expressing the HPV-16 E6 protein in their epidermis (K14E6WT) develop epithelial hyperplasia and squamous carcinomas. Numerous cellular proteins interact with E6, some of which can be grouped based on common amino acid motifs in their E6-binding domains. One such group, the PDZ partners, including hDLG, hSCRIBBLE, MUPP1, and MAGI, bind to the carboxy-terminal four amino acids of E6 through their PDZ domains. E6's interaction with the PDZ partners leads to their degradation. Additionally, E6's binding to PDZ proteins has been correlated with its ability to transform baby rat kidney cells in tissue culture and to confer tumorigenicity onto cells in xenograft experiments. To address whether the ability of E6 to bind PDZ domain partners is necessary for E6 to confer epithelial hyperproliferation in vivo, we generated transgenic mice that express in stratified squamous epithelia a mutant of E6 lacking the last six amino acids at its carboxyl terminus, E6Δ146-151, from the human keratin 14 (K14) promoter. The K14E6Δ146-151 mice exhibit a radiation response similar to that of the K14E6WT mice, demonstrating that this protein, as predicted, retains an ability to inactivate p53. However, the K14E6Δ146-151 mice fail to display epithelial hyperplasia. These results indicate that an interaction of E6 with PDZ partners is necessary for its induction of epithelial hyperplasia.

scholarly journals High-Risk Human Papillomavirus E6 Oncoproteins Interact with 14-3-3ζ in a PDZ Binding Motif-Dependent Manner

2012 ◽  
Vol 87 (3) ◽  
pp. 1586-1595 ◽  
Author(s):  
Siaw Shi Boon ◽  
Lawrence Banks
Keyword(s):  
Cervical Cancer ◽  
Human Papillomavirus ◽  
High Risk ◽  
Pdz Domain ◽  
Binding Motif ◽  
Dependent Manner ◽  
Hpv E6 ◽  
Hpv Types ◽  
High Risk Hpv ◽  
Hpv 18

ABSTRACTCervical cancer develops through the combined activities of the human papillomavirus (HPV) E6 and E7 oncoproteins. A defining characteristic of E6 oncoproteins derived from cancer-causing HPV types is the presence of a PDZ binding motif (PBM) at the extreme carboxy terminus of the protein which is absent from E6 proteins derived from the so-called low-risk HPV types. Within this PBM is also a protein kinase A (PKA) phospho-acceptor site, which is thought to negatively regulate the association of E6 with its PDZ domain-containing substrates. We can now show that phosphorylation of E6 by PKA and/or AKT confers the ability to interact with 14-3-3ζ. The interaction is direct and specific for the high-risk HPV E6 oncoproteins, although there are significant differences in the efficiencies with which HPV-16, HPV-18, and HPV-31 E6 oncoproteins can associate with 14-3-3ζ; this correlates directly with their respective susceptibilities to phosphorylation by PKA and/or AKT. We demonstrate here that the interaction between E6 and 14-3-3ζ also requires integrity of the E6 PBM, and downregulation of 14-3-3ζ results in a marked reduction in the levels of HPV-18 E6 expression in HeLa cells. Using phospho-specific anti-E6 antibodies, we also demonstrate significant levels of E6 phosphorylationin vivo. These studies redefine the potential relevance of the E6 PBM in the development of cervical cancer, suggesting that interaction with 14-3-3ζ, as well as the more well-established interactions with PDZ domain-containing substrates, is likely to be responsible for the biological activities attributed to this region of the high-risk HPV E6 oncoproteins.

Sign in / Sign up

Export Citation Format

Share Document