FIGURE 1:

Generation and characterization of EH mutants in Pan1p, Ede1p, and End3p. (A) Sequence alignments of the EH domains in Pan1p, End3p, Ede1p, and mouse Eps15. Residues chosen for mutagenesis are highlighted in red. Residues conserved highly and moderately throughout the family are indicated in black and gray, respectively. (B) The end3^EH mutant was temperature sensitive for growth at 39°C. A dilution series of cells was plated on YPD medium and incubated at 25, 37, or 39°C. (C) Average lifetimes of Pan1-GFP and End3-GFP ± SD in wild-type and mutant cells. Data were taken from 2-min movies with a 1-s frame interval. n = 50 patches for each strain. **p < 0.001. (D) Quantification of fluorescence intensity (red) and distance from site of patch formation (blue) as a function of time for patches of indicated GFP-tagged proteins. Each curve represents data from one patch. Behavior of three independent patches was plotted for each strain. (E) Distribution of Ede1-GFP and Ede1^EH-GFP patch lifetimes. Movies were taken with a 1-s frame interval. n = 50 patches for each strain.