Figure 6.
Distinct suppression and biochemical activity of exocyst gain-of-function mutants that mimic activation by Sro7 or Rho GTPases. (A) GST-Snc2 or GST-Snc2 E79,E82 were immobilized on beads and used in a pull down assay with purified wild-type exocyst or exocyst containing Exo84-N12I, Exo84-L365W, or Exo84-E626V. Quantitation of the pull-down experiments with error bars representing SD are shown to the right as well as a Coomassie stain of GST-fusion Snc2 protein used. P values were obtained using a two-tailed Student’s t test. *, P < 0.05; **, P < 0.01; *** P < 0.001; ns = no significant difference. (B) GST-Snc2 or GST- Snc2 E79,E82 were immobilized on beads and used to bind exocyst complexes containing either wild-type subunits, Exo84-N12I, or Exo70-I114F gain-of-function alleles as performed in A. Quantitation of binding was conducted as described above and is shown on the right. Error bars represent SD; P values were obtained using a two-tailed Student’s t test with significance annotated as above. (C) Purified wild-type exocyst and exocyst containing Exo70-I114F or Exo84-E626V were bound to beads containing immobilized GST-Sec4-GTPγS or control GST-Sec4-GDP. Quantitation of binding was conducted as described above and is shown on the right. Error bars represent SD; P values were obtained using a two-tailed Student’s t test with significance annotated as above. (D) Wild-type EXO84, wild-type EXO70, or the gain-of-function mutants EXO84-N12I, EXO84-L365W, EXO84-E626V and EXO70-I114F, vector (CEN) and 2µSRO7 were transformed into the sec2-41 (left) and cdc42-6 (right) mutant strains. The growth of three independent transformants is shown for each mutant allele, under permissive and restrictive conditions. (E) Wild-type EXO84, EXO84-N12I, EXO84-L365W and EXO84-N12I,L365W and vector were transformed into the sec2-41(left) and cdc42-6(right) mutant strains. The growth of three independent transformants is shown at the permissive and restrictive conditions. Source data are available for this figure: SourceData F6.

Distinct suppression and biochemical activity of exocyst gain-of-function mutants that mimic activation by Sro7 or Rho GTPases. (A) GST-Snc2 or GST-Snc2 E79,E82 were immobilized on beads and used in a pull down assay with purified wild-type exocyst or exocyst containing Exo84-N12I, Exo84-L365W, or Exo84-E626V. Quantitation of the pull-down experiments with error bars representing SD are shown to the right as well as a Coomassie stain of GST-fusion Snc2 protein used. P values were obtained using a two-tailed Student’s t test. *, P < 0.05; **, P < 0.01; *** P < 0.001; ns = no significant difference. (B) GST-Snc2 or GST- Snc2 E79,E82 were immobilized on beads and used to bind exocyst complexes containing either wild-type subunits, Exo84-N12I, or Exo70-I114F gain-of-function alleles as performed in A. Quantitation of binding was conducted as described above and is shown on the right. Error bars represent SD; P values were obtained using a two-tailed Student’s t test with significance annotated as above. (C) Purified wild-type exocyst and exocyst containing Exo70-I114F or Exo84-E626V were bound to beads containing immobilized GST-Sec4-GTPγS or control GST-Sec4-GDP. Quantitation of binding was conducted as described above and is shown on the right. Error bars represent SD; P values were obtained using a two-tailed Student’s t test with significance annotated as above. (D) Wild-type EXO84, wild-type EXO70, or the gain-of-function mutants EXO84-N12I, EXO84-L365W, EXO84-E626V and EXO70-I114F, vector (CEN) and 2µSRO7 were transformed into the sec2-41 (left) and cdc42-6 (right) mutant strains. The growth of three independent transformants is shown for each mutant allele, under permissive and restrictive conditions. (E) Wild-type EXO84, EXO84-N12I, EXO84-L365W and EXO84-N12I,L365W and vector were transformed into the sec2-41(left) and cdc42-6(right) mutant strains. The growth of three independent transformants is shown at the permissive and restrictive conditions. Source data are available for this figure: SourceData F6.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal