CLASS

Alternative Splicing Activity

COPY!
Gene Structure
Eukaryotic genes are composed of interspersed exons and introns.
 Exons (expressed sequences) contain the coding regions of the protein.
 Introns (interspersed sequences) are removed after transcription.

Most genes have 7 or 8 exons, with an average length of 145 base pairs (bp). Introns have an average
length of 3365 bp.

Splicing
Following transcription, introns are removed from the RNA transcript by a process called splicing. Splicing
results in a joining of all the exons that make up the coding region of a protein. In alternative splicing, the
same gene is processed in two or more ways. When the gene is spliced, different exons may be included
or excluded from the final transcript. For example, the initial transcript shown below could be spliced to
produce several different mRNA products, two of which are shown below.

This same initial transcript could be spliced in many additional ways not shown above.

Scientist used to think that “one gene, one protein”. Now we see that many proteins can be made from one
gene. Humans have only 20,000 - 25,000 genes. Earlier predictions of gene number based on the number
of proteins humans were estimated to produce was 100,000. Vertebrates have an average of 3.2 different
final transcripts per gene. About 59% of our genes are spliced into more than one different mRNA.

In this activity you will use colored pens or pencils to draw the mRNA before and
after it has been spliced. Similar to the diagram shown above, you will use empty
boxes to represent introns and colored boxes to represent exons.
Antibodies are proteins produced by B-cells, a type of white blood cell. Antibodies help prevent and clear infection by
an invading pathogen (often a disease causing bacteria or viruses).

Prior to infection, B-cells produce antibody proteins that remain embedded in

the cell membrane. Each B cell produces a different antibody, which recognizes
a different potential pathogen or invader. This antibody is embedded in the cell
membrane and protrudes from the cell.

The antibodies on the membrane surface of B-cells act as a receptor for

“antigens” found on an invading microbe. An antigen is a protein on the invading
bacteria or virus that our body recognizes as being “foreign.” During infection,
antigens on the pathogen surface bind the antibody on the surface of the B-cell.
This activates the B cell.

The activated B-cell then starts producing antibodies without the membrane
anchor that are secreted outside the B-cell into the blood stream where they can
act in fighting infection.

The same pre-mRNA transcript can be alternatively spliced to produce two different forms of an antibody protein; either the form
that is anchored in the cell membrane before an infection or the form that is secreted from the B-cell during an infection.

The antibody gene contains 8 exons. The functions of the various exons encoded by the antibody transcript are listed below
 Exon1 – encodes a signal peptide, this peptide is removed after translation. This signal peptide helps direct the antibody
protein from the rough ER (where it is made) to the Golgi apparatus.
 Exon 2 – codes for a part of the antibody protein that recognizes a particular antigen
 Exons 3 – 6 encode the constant region of the antibody. This is the part of the antibody that is the same in different
antibodies recognizing different pathogens.
 Exons 7 – 8 – encode a transmembrane region of the protein that anchors the antibody in the B-cell membrane.

For the antibody mRNA, use the following colors to represent the various exons:
 Exon 1 – Red  Exon 4 – Green  Exon 7 – Pink
 Exon 2 – Orange  Exon 5 – Blue  Exon 8 – Black
 Exon 3 – Yellow  Exon 6 – Purple

1) Draw and title the complete pre-mRNA strand as it would have been transcribed from the antibody gene. Draw introns and
exon boxes as large or as small as you would like, however please be sure to separate each exon with an intron and remember
that introns are usually much larger than exons. Please use the correct color for each exon. Then, provide a description of the
function of each exon in the antibody transcript.

2) Pretend you are a spliceosome and construct the mRNAs described below by removing the introns and exons that will not be
included.

a) mRNA 1: In resting B-cells the antibody mRNA contains all 8 exons. Using the correct colors, draw a diagram representing
this mRNA. Be sure to give your diagram a title. Then, answer the following questions in complete sentences:

i) What is the function of the protein produced from mRNA 1?

ii) In what type of cell and when is this mRNA generated?

b) mRNA 2: Once the B-cell has bound an antigen, it wants to secrete antibodies against that antigen. Now the transcript is
spliced so that only exons 1-6 are included. Using the correct colors, draw a diagram representing this mRNA. Be sure to
give you diagram a title. Then, answer the following questions in complete sentences:

i) What is the function of the protein produced from mRNA 2?

ii) In what type of cell and when is this mRNA generated?