Supplementary Figure 10: Characterization of Cdc48 FFF mutants and Npl4 with mutations in the β-strand finger
From: Structure of the Cdc48 ATPase with its ubiquitin-binding cofactor Ufd1–Npl4
. a, The indicated Cdc48 variants were subjected to gel filtration to analyze their oligomeric states. The position of hexamers is indicated. Note that mutation of the central Phe residue (FAF and AAA mutants) affects the migration of Cdc48 in gel filtration. b, Unfolding of poly-ubiquitinated Eos with wild-type Cdc48 and the indicated mutant of the conserved Tyr in the β-strand finger of Npl4. Data are shown as mean ± SD of n=3 technical replicates. c, A npl4-1 temperature-sensitive S. cerevisiae strain was transformed with a plasmid encoding wild-type Npl4 or the indicated mutations in insert-2, spotted in serial dilution, and incubated at the indicated temperatures for two days (30 and 37ºC) or three days (25ºC). d, Insert-2, in which mutations were made, is shown in purple and the Tyr in the β-strand finger is indicated (C. thermophilum Y472 corresponds to S. cerevisiae Y445).
