Question

In: Biology

2. Let’s assume that you do find movement utilizing cytoplasmic streaming. However, a new question is...

2. Let’s assume that you do find movement utilizing cytoplasmic streaming. However, a new question is now brought up. Is this movement dependent on cytoskeletal elements and motor proteins and if so, which ones? Describe how you could test to see which cytoskeletal elements/motor proteins were responsible for the movement of your protein. What would your data look like? Note you can’t knockout any motor proteins or cytoskeletal elements. They are essential to life; if you knock them out, the cell dies within a day or so. You have to find another method. Be creative.

Solutions

Expert Solution

The proteins involved the cytoplasmic streaming are the microfilament protein Actin & the motor protein Myosin. This occurs in a fashion similar to the sliding filament theory of muscular contraction where myosin proteins slide on the actin filaments with the help of energy from ATP.

Experiment to test which cytoskeletal elements/motor proteins are involved:

  • Using recombinant DNA technology, the gene for actin filament is tagged with Green fluorescence protein (GFP) gene.
  • When this recombinant fused-protein is expressed, the GFP showing fluorescence will allow us to track the actin protein molecules (with a fluorescence microscope) moving inside the cell during cytoplasmic streaming
  • As a control experiment, in another cell-line, the same recombinant-protein is expressed but this cell-line is treated with Cytochalasin D (which arrests cytoplasmic streaming)
  • On observing the two cell-lines, we will see that the GFP-tagged-actin molecules in the cell-line treated with Cytochalasin D will not show any movement. On the other hand, the other cell-line which is not treated with Cytochalasin D will show cytoplasmic streaming and hence the movement of the actin filaments.
  • This proves that actins are the cytoskeletal proteins that are involved in cytoplasmic streaming in association with myosin motor proteins.
  • To test for myosin's involvement, a similar experiment can be carried out but this time myosin will be GFP-tagged

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