Please answer all parts, this is all one question. mutant 1 1. To investigate th
ID: 189641 • Letter: P
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Please answer all parts, this is all one question.
mutant 1 1. To investigate the behavior of cofilin, both in vivo and in vitro experiments were performed with wild-type cofilin and two mutants. Mutant 1 has two charged amino acids replaced with alanines. Mutant 2 has three changed amino acids replaced with alanines. For the in vivo experiment, cultures of yeast cells were transfected with one of the three genes. Yeast cells have "actin patches", regions dense with microfilaments associated with the cell cortex. The transfected cultures were treated with latrunculin, a drug that sequesters free globular actin, at time zero. Then the cells were examined under an optical microscope. (50 points total) Cells with actin patches (3%) mutant 2 wild type Time (min) a. What is the purpose of treating the cultures with latrunculin? (10 points) control mutant 1 Light Scattering (400 nm) b. What is the action of cofilin? (10 points) mutant 2 30 wild type Time (min) c. How do the mutants differ from wild-type? (10 points)Explanation / Answer
1 A) latruculin is a drug that sequestered the monomeric G-filaments and prevents actin polymerization and thus ceases the lemellepodium movement and leads to the destruction of microfilament. latruculin used in culture to see the effect of this drug on mutant species and to determine the advantage of mutation over wild type.
1 B) Cofilin is deeply involved in the regulation of actin assembly in developing skeletal muscle. In this study, to examine further the function of cofilin in living myogenic cells in culture and in mutant.
1 C) Mutants are more resistant towards the latruculin drug than wild type and among mutants the mutant 1 shows more resistant that mutant 2 which determines that replacement of 2 charged aminoacid with alanine is the advantageous prospetive for making yeast cells resistive to these drug and less regulated by cofilin.
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