Generation and evaluation of a human corneal model cell system for ophthalmologic issues using the HPV16 E6/E7 oncogenes as uniform immortalization platform

2013 ◽  
Vol 85 (4-5) ◽  
pp. 161-172 ◽  
Author(s):  
Simon Schulz ◽  
Thorsten Steinberg ◽  
David Beck ◽  
Pascal Tomakidi ◽  
Rosita Accardi ◽  
...  
Keyword(s):  
Cell System ◽  
Hpv16 E6 ◽  
Model Cell

Spheroid organization kinetics of H35 rat hepatoma model cell system on elastin-like polypeptide-polyethyleneimine copolymer substrates

2013 ◽  
Vol 102 (3) ◽  
pp. 852-861 ◽  
Author(s):  
Paul A. Turner ◽  
C. Andrew Weeks ◽  
Austin J. McMurphy ◽  
Amol V. Janorkar
Keyword(s):  
Cell System ◽  
Model Cell ◽  
Kinetics Of ◽  
Hepatoma Model ◽  
Rat Hepatoma

scholarly journals The Extreme C-Terminal Region of Gαs Differentially Couples to the Luteinizing Hormone and β2-Adrenergic Receptors

2011 ◽  
Vol 25 (8) ◽  
pp. 1416-1430 ◽  
Author(s):  
Geneva DeMars ◽  
Francesca Fanelli ◽  
David Puett
Keyword(s):  
G Protein ◽  
Comparative Modeling ◽  
Cell System ◽  
Amino Acid Residues ◽  
Intracellular Loop ◽  
C Terminus ◽  
Model Cell ◽  
Rigid Body Docking ◽  
Molecular Landscape ◽  
Dynamics Simulations

The mechanisms of G protein coupling to G protein-coupled receptors (GPCR) share general characteristics but may exhibit specific interactions unique for each GPCR/G protein partnership. The extreme C terminus (CT) of G protein α-subunits has been shown to be important for association with GPCR. Hypothesizing that the extreme CT of Gαs is an essential component of the molecular landscape of the GPCR, human LH receptor (LHR), and β2-adrenergic receptor (β2-AR), a model cell system was created for the expression and manipulation of Gαs subunits in LHR+ s49 ck cells that lack endogenous Gαs. On the basis of studies involving truncations, mutations, and chain extensions of Gαs, the CT was found to be necessary for LHR and β2-AR signaling. Some general similarities were found for the responses of the two receptors, but significant differences were also noted. Computational modeling was performed with a combination of comparative modeling, molecular dynamics simulations, and rigid body docking. The resulting models, focused on the Gαs CT, are supported by the experimental observations and are characterized by the interaction of the four extreme CT amino acid residues of Gαs with residues in LHR and β2-AR helix 3, (including R of the DRY motif), helix 6, and intracellular loop 2. This portion of Gαs recognizes the same regions of the two GPCR, although with differences in the details of selected interactions. The predicted longer cytosolic extensions of helices 5 and 6 of β2-AR are expected to contribute significantly to differences in Gαs recognition by the two receptors.

scholarly journals FOXM1 drives HPV+ HNSCC sensitivity to WEE1 inhibition

2020 ◽  
Vol 117 (45) ◽  
pp. 28287-28296
Author(s):  
Ahmed Diab ◽  
Hakan Gem ◽  
Jherek Swanger ◽  
Hee Yeon Kim ◽  
Kaleb Smith ◽  
...  
Keyword(s):  
Kinase Inhibitor ◽  
Single Agent ◽  
Genomic Analysis ◽  
Clinical Problem ◽  
Hpv Infection ◽  
Cell System ◽  
Cell Cycle Checkpoints ◽  
The Cancer Genome Atlas ◽  
Primary Tumors ◽  
Hpv16 E6

Head and neck squamous cell carcinoma (HNSCC) associated with high-risk human papilloma virus (HPV) infection is a growing clinical problem. The WEE1 kinase inhibitor AZD1775 (WEE1i) overrides cell cycle checkpoints and is being studied in HNSCC regimens. We show that the HPV16 E6/E7 oncoproteins sensitize HNSCC cells to single-agent WEE1i treatment through activation of a FOXM1-CDK1 circuit that drives mitotic gene expression and DNA damage. An isogenic cell system indicated that E6 largely accounts for these phenotypes in ways that extend beyond p53 inactivation. A targeted genomic analysis implicated FOXM1 signaling downstream of E6/E7 expression and analyses of primary tumors and The Cancer Genome Atlas (TCGA) data revealed an activated FOXM1-directed promitotic transcriptional signature in HPV+ versus HPV- HNSCCs. Finally, we demonstrate the causality of FOXM1 in driving WEE1i sensitivity. These data suggest that elevated basal FOXM1 activity predisposes HPV+ HNSCC to WEE1i-induced toxicity and provide mechanistic insights into WEE1i and HPV+ HNSCC therapies.

The effects of desmethylimipramine on cyclic AMP-stimulated gene transcription in a model cell system

2005 ◽  
Vol 70 (5) ◽  
pp. 762-769 ◽  
Author(s):  
J.K. Richards ◽  
W. Abdel-Razaq ◽  
T.E. Bates ◽  
D.A. Kendall
Keyword(s):  
Cyclic Amp ◽  
Gene Transcription ◽  
Cell System ◽  
Model Cell

Use of single-cell imaging techniques to assess the regulation of cAMP dynamics

2006 ◽  
Vol 34 (4) ◽  
pp. 468-471 ◽  
Author(s):  
D. Willoughby ◽  
D.M.F. Cooper
Keyword(s):  
Single Cell ◽  
Real Time ◽  
Imaging Techniques ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Cell System ◽  
Cellular Processes ◽  
Signalling Molecule ◽  
Anchoring Protein ◽  
Model Cell

cAMP is a ubiquitous intracellular signalling molecule that can regulate a wide array of cellular processes. The diversity of action of this second messenger owes much to the localized generation, action and hydrolysis of cAMP within discrete subcellular regions. Further signalling specificity can be achieved by the ability of cells to modulate the frequency or incidence of such cAMP signals. Here, we discuss the use of two cAMP biosensors that measure real-time cAMP changes in the single cell, to address the mechanisms underlying the generation of dynamic cAMP signals. The first method monitors sub-plasmalemmal cAMP changes using mutant cyclic nucleotide-gated channels and identifies an AKAP (A-kinase-anchoring protein)–protein kinase A–PDE4 (phosphodiesterase-4) signalling complex that is central to the generation of dynamic cAMP transients in this region of the cell. The second study uses a fluorescence resonance energy transfer-based cAMP probe, based on Epac1 (exchange protein directly activated by cAMP 1), to examine interplay between Ca2+ and cAMP signals. This study demonstrates real-time oscillations in cAMP driven by a Ca2+-stimulated AC (adenylate cyclase) (AC8) and subsequent PDE4 activity. These studies, using two very different single-cell cAMP probes, broaden our understanding of the specific spatiotemporal characteristics of agonist-evoked cAMP signals in a model cell system.

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