Equilibrium dissociation and unfolding of human papillomavirus E2 transactivation domain

Singh, Nitu; Kanthaje, Shruthi; Bose, Kakoli

doi:10.1016/j.bbrc.2015.05.057

Cited by 1 publication

(2 citation statements)

References 32 publications

(45 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Later, MBP-tagged DED construct was induced with 0.4 mM IPTG at optical density (OD 600 ) = 0.6 and were grown overnight at 18 °C. For isolation of the complex, the cell lysate in buffer-A (20 mM Na 2 HPO 4 /NaH 2 PO 4 , pH 7.8, 250 mM NaCl, 0.1 mM DTT, and 1% glycerol) was passed through pre-equilibrated Ni-IDA or amylose resin and purified as described earlier 36 . The complexes were separated on 12% SDS-PAGE and immunoblotted with 1:1000 dilution of anti-E2 antibody (sc-26939, Santa Cruz Biotechnology, Inc.).…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Insights into the mechanism of human papillomavirus E2-induced procaspase-8 activation and cell death

Singh

Senapati

Bose

2016

Sci Rep

Self Cite

View full text Add to dashboard Cite

High-risk human papillomavirus (HR-HPV) E2 protein, the master regulator of viral life cycle, induces apoptosis of host cell that is independent of its virus-associated regulatory functions. E2 protein of HR-HPV18 has been found to be involved in novel FADD-independent activation of caspase-8, however, the molecular basis of this unique non-death-fold E2-mediated apoptosis is poorly understood. Here, with an interdisciplinary approach that involves in silico, mutational, biochemical and biophysical probes, we dissected and characterized the E2-procasapse-8 binding interface. Our data demonstrate direct non-homotypic interaction of HPV18 E2 transactivation domain (TAD) with α2/α5 helices of procaspase-8 death effector domain-B (DED-B). The observed interaction mimics the homotypic DED-DED complexes, wherein the conserved hydrophobic motif of procaspase-8 DED-B (F122/L123) occupies a groove between α2/α3 helices of E2 TAD. This interaction possibly drives DED oligomerization leading to caspase-8 activation and subsequent cell death. Furthermore, our data establish a model for E2-induced apoptosis in HR-HPV types and provide important clues for designing E2 analogs that might modulate procaspase-8 activation and hence apoptosis.

show abstract

Section: Methodsmentioning

confidence: 99%

“…Structures of procaspase-8 DED-AB (1–190) and HPV18 E2 TAD were homology-modeled as described previously 22 36 . E2 TAD was docked on to procaspase-8 DED using HADDOCK 37 .…”

Section: Methodsmentioning

confidence: 99%