You study a serine/threonine protein kinase, which is likely involved in the regulation of actin dynamics....

You study a serine/threonine protein kinase, which is likely involved in the regulation of actin dynamics. You

have good knowledge about the kinase (such as its kinase domain information and which residues are

catalytic sites etc). You found a RhoGEF by yeast two-hybrid screening using the kinase as bait. Further,

through co-immunoprecipitation assay, you confirmed that the kinase specifically interacts with the

RhoGEF. Since RhoGEF is well known to regulate actin oligomerization, you expect that the protein might

provide an important clue about the role of the kinase in the regulation of actin dynamics. How would you

define a signaling pathway from the kinase to actin polymerization via the RhoGEF as a mediator? What

would be your questions to define the pathway, and what experiments would you do to address them?

Expert Solution

The interaction between the serine-threonine kinase and the RhoGEF has been established by the above experiments. But what type of interaction is this and what does it do on the expression levels is the next question. Whether this interaction increases or decreases the level of RhoGEF in the cell and if it increases what sort of other pathways are upregulated and what sort of other pathways are downregulated. With regard to the actin dynamics, RhoGEF helps in the oligomerization of the actin. The protein serine-threonine kinase can upregulate the RhoGEF and thereby increase the actin oligomerization for the movement of the molecules within the cell or else they may downregulate the RhoGEF to stop the actin oligomerization and halt the movement of molecules within the cell. To know about this expression levels, qRT-PCR studies can be performed and the expression levels at various time intervals can be studied or at various hormonal changes inside the cell to know which pathway this is affecting.

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gladiator answered 1 year ago

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