Question

It is suggested that BIP1 is a kind of BZR1-interacting protein. Please design a research to study the BIP1 function in brassinosteroid signaling. And hence how it affect the growth of the plant.

Answer 1

The following techniques could be used to know the function of BIP1 in the signalling pathway:

1. Create a mutant BIP1 and a wild type BIP1 clones and express them in cells. There will be changes in the cells.

2. Do the yeast 2 hybrid assay again with mutant BIP1 and check for its interaction. Mostly it shouldnt be interacting.

3. Because the mutant BIP1 is not interacting with the BZR1, the pathway will accumulate some proteins in it and also the mRNA levels will vary. Hence do a mRNA and proteomic analysis of the wild type and the mutant cells.

4. This will give us an insight into which protein level is getting overexpressed and which one is downregulated and also same goes for the mRNA.

5. Once this is done, you can hypothesise that the BZR1 is a nuclear protein and is inactive in phosphorylated state. Do a western blot with the anti-phosph BZR1 antibodies for both mutant and wild type BIP1 expressing cells. In mutant if the band is seen then BIP1 is the protein required for the activation of the BZR1 protein and if band is seen in the wild type one, then the BIP1 is a protein that inactivates the BZR1 protein.

Thus by doing these experiments one can confirm the functions of BIP1 in the cells BZR1 pathway.

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