Take Home Exam Problem

In recent years, several cancer-derived lines of cells have been used in order to better
understand cell differentiation and development. The HT-29-M6 are one type of cells that are
obtained from cancer cells in the epithelium of human intestines. With these cells, the role of E2
ubiquitin-conjugation enzymes, such variants of these proteins are designated as UEV proteins.
UEV-1 is an inactive variant of one of the E2 complex proteins. From the graph in Figure 11A
(pg. 585), no significant change in the number of cells produced occurs, but the graph in Figure
11B shows drastic increases in DNA replication at 3 hrs. and about 21 hrs.

a.) The peak intensity of the counts per minute (cpm) in Figure 11B is about 0.120 cpm.
Convert this value to microcuries (Ci) and also to becquerels (Bq). (Hint: 1Ci = 2.22 x 1012
cpm = 3.7 x 1010 Bq).

b.) Explain how the DNA replication rate can go up without increasing the cell count at that
point in the experiment. Explain what this UEV protein variation may do to keep the cell count
relatively constant, yet cause the DNA replicate more rapidly.

c.) Normal human cells are termed diploid because they have 2 copies of most of the
chromosomes. What is the name for cells containing 3, 4, and more than 4 copies of the
genome?

d.) Cancerous cells are ones that lack control over growth and function of the cells. Cells
that contain multiple copies of the genes may lead to growths of cancer. How do cells with
differing copy numbers of the genome cause a loss in control of protein function?

Answers
a.) 0.120 cpm x 1 Ci x 1000000 Ci = 5.41 x 10-11 Ci
12
2.22 x 10 cpm 1 Ci

5.41 x 10-14 Ci x 3.7 x 1010 Bq = 0.00200 Bq

1 Ci

b.) In order for the DNA replication rate to increase without an increase in cell count, the
cells must be performing replication without mitotically dividing. The UEV-1 protein variation
must effect the cells' ability to signal mitotic division of the membrane along with an increase in
the rate of DNA replication. Many of these mutant cells contain multiple nucleuses, which
explains how the DNA replication rate can increase without changing the cell count significantly.

c.) 3 is triploid, 4 is tetraploid, and many is polyploid.

d.) In polyploids, the genes of the chromosomes are transcribed and translated at a higher
rate because there are more copies of the gene. This causes an increase in the amount of proteins
produced by the cell, which is usually not a problem for the cell. This is because all of the genes
are transcribed and translated at the same rate. Therefore, there is a certain enzyme balance
obtained by having multiple copies of the entire genome. This term shows that the relative
amounts of proteins in polyploid are the same as for diploid cells.
 This increased amount of proteins does cost the cell more energy and more time to
complete replication, transcription, and translation. Polyploidy is not well tolerated in animals,
humans specifically, because cell differentiation is very dependent upon appropriate timing of
cell divisions. With those extra copies of the chromosomes, the cell divisions and
differentiations occur before the cell has completed its tasks. This causes some of the cells to
have incomplete or improper cell divisions, which leads to uncontrolled growth of some cells
and an absence of other important cell types. This gives the organism a lack of cell
differentiation and a loss of the functions of that type of cell.