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Cellular interpretation of the long-range gradient of Four-jointed activity in the Drosophila wing.

eLife (2015-02-25)
Rosalind Hale, Amy L Brittle, Katherine H Fisher, Nicholas A M Monk, David Strutt
ABSTRACT

To understand how long-range patterning gradients are interpreted at the cellular level, we investigate how a gradient of expression of the Four-jointed kinase specifies planar polarised distributions of the cadherins Fat and Dachsous in the Drosophila wing. We use computational modelling to test different scenarios for how Four-jointed might act and test the model predictions by employing fluorescence recovery after photobleaching as an in vivo assay to measure the influence of Four-jointed on Fat-Dachsous binding. We demonstrate that in vivo, Four-jointed acts both on Fat to promote its binding to Dachsous and on Dachsous to inhibit its binding to Fat, with a bias towards a stronger effect on Fat. Overall, we show that opposing gradients of Fat and Dachsous phosphorylation are sufficient to explain the observed pattern of Fat-Dachsous binding and planar polarisation across the wing, and thus demonstrate the mechanism by which a long-range gradient is interpreted.

MATERIALS
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Product Description

Sigma-Aldrich
Shields and Sang M3 Insect Medium, With L-glutamine and potassium bicarbonate., liquid, sterile-filtered, suitable for insect cell culture