Structure of the intact transactivation domain of the human papillomavirus E2 protein

Antson, A.A.; Burns, Julie E.; Moroz, Olga V.; Scott, David J.; Sanders, Cyril M.; Bronstein, Igor B.; Dodson, Guy; Wilson, K.S.; Maitland, Norman J.

doi:10.1038/35001638

Cited by 99 publications

(121 citation statements)

References 23 publications

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“…We therefore studied the relationship between ectopically coexpressed Adaptor-independent activation of caspase 8HPV18 E2 protein induces caspase 8 oligomerization within filaments independently of FADD through direct interaction. Analysis of the crystal structure of the aminoterminal domain of the HPV16 E2 protein did not reveal the presence of an obvious DFD typically composed of six a-helices structures, 14 indicating that E2 may not act as a typical DFD adaptor to induce caspase 8 activation. We therefore investigated whether caspase oligomerization within filaments in the presence of E2 could be mediated by the endogenous DFD adaptor FADD, which should then be found in the insoluble filaments together with E2 and caspase 8.…”

Section: Resultsmentioning

confidence: 99%

“…7,25 RFP-E2 was obtained by inserting the RFP ORF (from pDSRed2) in place of the GFP ORF in the GFP-E2 expression plasmid. E2 helices H1 (aa 1-26), H2 (aa 27-53) or H3 (aa 54-88) corresponding to the three first a-helices of the amino terminal of HPV18 E2 as determined by the crystal structure, 14 were cloned by a conventional PCR-based technique in frame with GFP. The caspase 8 prodomain (aa 1-181) was cloned in pcDNA, DED A (aa 1-93) DED B (aa 94-181) were cloned in frame with GFP and caspase domain CD (aa180-480) in frame with a N-terminal FLAG tag in pcDNA 3.…”

Section: Methodsmentioning

confidence: 99%

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Direct activation of caspase 8 by the proapoptotic E2 protein of HPV18 independent of adaptor proteins

Thierry

Demeret

2008

Cell Death Differ

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The self-activation of initiator caspases is dependent on their oligomerization driven by interaction with the death fold domains (DFD) of adaptor proteins. Here, we show that the E2 protein of human papillomavirus type 18 triggers apoptosis by assembling cytoplasmic filaments together with caspase 8, in which its efficient self-activation occurs. The E2 protein binds directly to the death effector domains (DED) of caspase 8 through non-DFD interaction. This interaction is independent of FADD, but it can cooperate with FADD homotypic binding to caspase 8 to induce its oligomerization; hence cell death, while it is antagonized by competitive binding of MC159 FLICE inhibitory protein. The amino-terminal domain of E2 contains a 27 amino-acid a-helix, which is necessary and sufficient to induce caspase oligomerization and cell death. Our results provide evidence for adaptorindependent oligomerization of caspase 8, mediated by non-DFD direct interactions with the HPV18 E2 protein, thus deciphering a new pathway for caspase 8 activation.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Direct activation of caspase 8 by the proapoptotic E2 protein of HPV18 independent of adaptor proteins

Thierry

Demeret

2008

Cell Death Differ

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“…Crystal structures have been reported for the TAD of HPV16 (amino acids 1-201) (20) and for a truncated TAD of HPV18 (amino acids 66 -215) lacking two N-terminal ␣-helices (21). The HPV16 and -18 TADs share ϳ45% sequence identity (see Fig.…”

mentioning

confidence: 99%

Crystal Structure of the E2 Transactivation Domain of Human Papillomavirus Type 11 Bound to a Protein Interaction Inhibitor

Wang

Coulombe²,

Cameron

et al. 2004

Journal of Biological Chemistry

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Interaction between the E2 protein and E1 helicase of human papillomaviruses (HPVs) is essential for the initiation of viral DNA replication. We recently described a series of small molecules that bind to the N-terminal transactivation domain (TAD) of HPV type 11 E2 and inhibits its interaction with E1 in vitro and in cellular assays. Here we report the crystal structures of both the HPV11 TAD and of a complex between this domain and an inhibitor, at 2.5-and 2.4-Å resolution, respectively. The HPV11 TAD structure is very similar to that of the analogous domain of HPV16. Inhibitor binding caused no significant alteration of the protein backbone, but movements of several amino acid side chains at the binding site, in particular those of Tyr-19, His-32, Leu-94, and Glu-100, resulted in the formation of a deep hydrophobic pocket that accommodates the indandione moiety of the inhibitor. Mutational analysis provides functional evidence for specific interactions between Tyr-19 and E1 and between His-32 and the inhibitor. A second inhibitor molecule is also present at the binding pocket. Although evidence is presented that this second molecule makes only weak interactions with the protein and is likely an artifact of crystallization, its presence defines additional regions of the binding pocket that could be exploited to design more potent inhibitors.Human papillomaviruses (HPVs) 1 are the etiological agents of malignant and benign lesions of the differentiating squamous or mucosal epithelium, notably of cervical cancer. Approximately 25 HPV types replicate in mucosal tissues of the anogenital tract. HPV16, -18, and -31 are the most prevalent "high-risk" types found in pre-cancerous or malignant lesions of the cervix. HPV6 and -11 are the most common "low-risk" types, which cause benign genital warts (condyloma acuminata), a less serious condition but one of the most common sexually transmitted diseases (1). Currently, no specific antivirals are available for the treatment of HPV infections.The small circular double-stranded DNA genome of papillomavirus is actively maintained as a multicopy episome in the nucleus of infected epithelial cells. This process is dependent on replication of the viral genome by the viral E1 and E2 proteins, in conjunction with the host DNA replication machinery. E2 is a sequence-specific DNA-binding protein that has a number of functions in the viral lifecycle. In addition to its role in the initiation of viral DNA replication, E2 is involved in regulating the transcription of viral genes (2-7), and in the segregation of the viral genome during cell division (8, 9). As a replication initiation factor, E2 binds with high affinity to specific sites located within the viral origin (ori) to help recruit it to the E1 helicase (10 -13). Formation of a ternary complex between E1, E2, and the origin relies not only on the interaction of E1 and E2 with specific DNA sequences at the origin but is also critically dependent on a direct interaction between these two proteins (14 -18).The 40-kDa E2 prote...

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“…The results of transcription repression studies reported here support an antiparallel DNA loop in which the GalR dimer bound to O I stacks upon the one bound to O E , consistent with the previous theoretical calculations (Geanacopoulos et al 2001). The approach reported here can be generally applicable to trace DNA trajectories in DNA loop-containing multiprotein complexes based on the "modular" principle (Antson et al 2000;Dodd et al 2001).…”

mentioning

confidence: 99%

DNA trajectory in the Gal repressosome

Semsey¹,

Tolstorukov²,

Virnik³

et al. 2004

Genes Dev.

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Structure of the intact transactivation domain of the human papillomavirus E2 protein

Cited by 99 publications

References 23 publications

Direct activation of caspase 8 by the proapoptotic E2 protein of HPV18 independent of adaptor proteins

Direct activation of caspase 8 by the proapoptotic E2 protein of HPV18 independent of adaptor proteins

Crystal Structure of the E2 Transactivation Domain of Human Papillomavirus Type 11 Bound to a Protein Interaction Inhibitor

DNA trajectory in the Gal repressosome

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