27 Jun BIO 320 Homework 2
Question
SNAREs exist as complementary partners that carry out membrane fusions between appropriate vesicles and their target membranes. In this way, a vesicle with a particular variety of v-SNARE will fuse only with a membrane that carries the complementary t-SNARE. In some instances, however, fusions of identical membranes (homotypic fusions) are known to occur. For example, when a yeast cell forms a bud, vesicles derived from the mother cell’s vacuole move into the bud where they fuse with one another to form a new vacuole. These vesicles carry both v-SNAREs and t-SNAREs. You are wondering whether both types of SNAREs are essential for this homotypic fusion event.
To test this point, you have developed an ingenious assay for fusion of vacuolar vesicles. You prepare vesicles from two different mutant strains of yeast: strain B has a defective gene for vacuolar alkaline phosphatase (Pase); strain A is defective for the protease that converts the precursor of alkaline phosphatase (pro-Pase) into its active form (Pase) (Figure A). Neither strain has active alkaline phosphatase, but when extracts of the strains are mixed, vesicle fusion generates active alkaline phosphatase, which can be easily measured.
Now you delete the genes for the vacuolar v-SNARE, t-SNARE, or both in each of the two yeast strains. You prepare vacuolar vesicles from each and test them for their ability to fuse, as measured by the alkaline phosphatase assay (Figure B).
(A) By comparing experiment 1 with other 10 experiments, what can you conclude?
(B) By comparing experiments (3,4,6,7,8,9) together with experiments (2,5,10,11), what can you conclude?
(C) Does it matter which kind of SNARE is on which vesicle, and by what comparisons do you get this conclusion?
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