Fig. 3
From: Cohesin cleavage by separase is enhanced by a substrate motif distinct from the cleavage site
The LPE motif is important for cleavage of a separase biosensor in vivo. a Schematic of the separase biosensor used to evaluate cleavage in vivo, which includes histone H2B, red fluorescent protein (RFP), the indicated Scc1 fragment, and green fluorescent protein (GFP)33. b Time course of wild-type (WT) biosensor cleavage by separase, showing green fluorescence (left), red fluorescence (center), and merged images (right). Time zero is the last time point before the onset of chromosome segregation. Biosensor cleavage is indicated by reduced green fluorescence relative to red fluorescence. Scale bar: 20 μM. c Representative images showing late anaphase fluorescence of biosensor variants carrying mutations in Scc1 (WT, wild-type, copied from a; NC, non-cleavable mutations at sites 1 and 2; LP → AA, mutations of 255LP; ∆10aa, deletion of aa 251 to 260, which contain the LPE motif). Scale bars: 20 μM. d Quantification of the loss of GFP fluorescence in the four biosensor variants shown in c. Data points indicate means (±SD) from between 15 and 30 cells. Source data are provided in the Source Data file
