Ablation of Whirlin Long Isoform Disrupts the USH2 Protein Complex and Causes Vision and Hearing Loss (original) (raw)
Figure 4
Whirlin directly interacts with usherin.
(A) A yeast two-hybrid analysis demonstrates that mouse whirlin, through its N-terminal two PDZ domains (whirlin N-term), interacts with the C-terminal fragment of mouse usherin. The “+” sign denotes growth of large, white yeast colonies and “−” denotes absence of colonies. This experiment was conducted in two orientations. (B) A GST pull-down assay demonstrates that the intact C-terminal fragments of mouse and frog usherin, but not the mutant ones, which lack the PDZ-binding motif, were able to pull down the endogenous whirlin from the retinal lysate, indicating that the PDZ-binding motif of usherin is involved in the interaction between whirlin and usherin. The arrows on the GST blot indicate the positions of mouse and frog GST-fused usherin fragments and GST. The multiple bands below the mouse and frog GST-fused usherin fragments are their degraded products.