Help:Tutorial:Step 6

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Target genes are activated by the E-box element

After the Clock/Bmal1 dimer binds to the E-box element, transcription of several target genes are activated. This simplified version of the Circadian clock pathway, will restrict to the mPer/mCry genes, which will act as a negative feedback loop. As long as Per1 or Per2 exist as monomers, they will be phosphorylated by casein kinase 1 delta or epsilon, followed by degradation. In the pathway drawing we simplify it a bit by leaving out the phosphorylation event, resulting in the following representation:

transcription_degradation.png
Drawing the transcription of mPer/mCry
and degradation of mPer

1. Stack the mCry/mPer DataNodes

To illustrate that the mCry/mPer genes are activated by the E-box element, we can stack them below the 'Gene' label. Place the mPer/mCry DataNodes below the 'Gene' label and stack them vertically, using the stackverticalcenter.gif stack vertically button.

2. Copy the gene-products

The genes will be translated into proteins, to show this on the pathway, copy the stacked genes and move them to the right of the stacked DataNodes. Separate the mPer DataNodes from the mCry DataNodes to show that they exist as seperate proteins.

3. Group the gens and gene-products

In this example we will use nested groups (groups within other groups), so the order of grouping and linking is important. We can create the following groups:

  1. the Per1/2 and Cry1/2 genes under the Gene label
  2. group of Per and Cry pairs
    1. the Per1/2 gene-products
    2. the Cry1/2 gene-products

The tutorial video shows the order of grouping for group 2. Start with creating the lowest level groups (group 2.1 and 2.2) and then group these to form group 2.

4. Draw a line from the genes to the gene-products

To illustrate that the genes are translated into proteins, we draw a dashed arrow from the stacked genes to the copied genes. Connect the start point to group 1, connect the end point to group 2.

5. Create a degradation shape

As long as Per1 or Per2 is not dimerized with Cry1 or Cry2, it will be degraded. We can indicate degradation by using the degradation shape:

degradation.png

You will notice that this shape is not on the toolbar, so to insert it, we have to insert another shape (e.g. a rectangle) and change it's Shape Type property. Insert a rectangle, select it and change the Shape Type property in the properties table to mim-degradation:

shapetype.png

When you selected mim-degradation as property, you will see the rectangle change into a degradation shape.

6. Draw an arrow from the mPer DataNodes to the degradation shape

To show that the mPer proteins are degraded, draw an dashed arrow from the two mPer DataNodes to the degradation shape. Connect the start of this line to group 2.1 to specify that only the mPer proteins are degraded.

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