Figure 1. C-terminal KA-1 domain and basic motif mediate Cdr2 membrane binding. (A) Schematic representation of full-length Cdr2 (top) and Cdr2 C-terminal cortex-anchoring domain (Cdr2-Cter, residues 591–747; bottom) composed of a basic region (blue) and a KA-1 domain (green on top and rainbow on the bottom). Predicted α helices and β sheets of the KA-1 domain are shown. (B) Structural model of Cdr2 KA-1 domain derived from the S. cerevisiae Kcc4 KA-1 crystal structure (Protein Data Bank accession no. 3OST). Seven positively charged residues sticking out of the domain are highlighted in blue. (C) Epifluorescence medial plane images of mEGFP-tagged Cdr2 mutants of the positively charged residues of Cdr2 KA-1 shown in B (Cdr2RR*: R624Q and R628Q; Cdr2RKRKR*: R682Q, K684N, R685Q, K692N, and R695Q; Cdr2RR*-RKRKR*: combination of all mutations), in the basic region upstream of Cdr2 KA-1 (Cdr2bsc*: K598N, H599Q, R600Q, R601Q, R602Q, K612N, K613N, and K614N), or in both (Cdr2bsc*-RKRKR*). Bar, 5 µm. (D) Mean FRAP of mEGFP-tagged Cdr2 or Cdr2RR* and Cdr2Δbsc mutants on the medial cortex as indicated. Error bars show SDs. Cdr2 t1/2 = ∼3 min; Cdr2RR* t1/2 = ∼3 min; Cdr2bscΔ t1/2 < 1 min. B, bleach.
Figure 1.

C-terminal KA-1 domain and basic motif mediate Cdr2 membrane binding. (A) Schematic representation of full-length Cdr2 (top) and Cdr2 C-terminal cortex-anchoring domain (Cdr2-Cter, residues 591–747; bottom) composed of a basic region (blue) and a KA-1 domain (green on top and rainbow on the bottom). Predicted α helices and β sheets of the KA-1 domain are shown. (B) Structural model of Cdr2 KA-1 domain derived from the S. cerevisiae Kcc4 KA-1 crystal structure (Protein Data Bank accession no. 3OST). Seven positively charged residues sticking out of the domain are highlighted in blue. (C) Epifluorescence medial plane images of mEGFP-tagged Cdr2 mutants of the positively charged residues of Cdr2 KA-1 shown in B (Cdr2RR*: R624Q and R628Q; Cdr2RKRKR*: R682Q, K684N, R685Q, K692N, and R695Q; Cdr2RR*-RKRKR*: combination of all mutations), in the basic region upstream of Cdr2 KA-1 (Cdr2bsc*: K598N, H599Q, R600Q, R601Q, R602Q, K612N, K613N, and K614N), or in both (Cdr2bsc*-RKRKR*). Bar, 5 µm. (D) Mean FRAP of mEGFP-tagged Cdr2 or Cdr2RR* and Cdr2Δbsc mutants on the medial cortex as indicated. Error bars show SDs. Cdr2 t1/2 = ∼3 min; Cdr2RR* t1/2 = ∼3 min; Cdr2bscΔ t1/2 < 1 min. B, bleach.

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