Exam 17: The Cell Cycle

In the following schematic drawing of a vertebrate prometaphase chromosome as seen under a microscope, indicate whether the chromosome is more likely to be closer to pole A or pole B. Write down A or B as your answer.

Indicate true (T) and false (F) statements below regarding cytokinesis in animal cells. Your answer would be a four-letter string composed of letters T and F only, e.g. TFFF. ( ) The force for cytokinesis is generated by kinesin motors on microtubule bundles that form the contractile ring. ( ) As the contractile ring constricts, its thickness increases to keep a constant volume. ( ) The midbody forms from bundles of actin and myosin II. ( ) Local activation of Ran GTPase triggers the assembly and contraction of the contractile ring.

The microtubule-binding protein Patronin binds to the minus ends of microtubules at the spindle pole and protects them from the effect of catastrophe factors. If Patronin activity is inhibited by injecting an anti-Patronin antibody into Drosophila melanogaster embryos prior to cellularization, anaphase B is suppressed and the spindles are significantly shorter. Which of the forces (1 or 2) in the following schematic diagram do you think is mostly responsible for anaphase B in this organism at this stage? Does the effect of Patronin inhibition resemble that of kinesin-5 inhibition (I) or overactivation (O)?

Indicate whether each of the following phosphorylation events typically activates (A) or inactivates (I) the protein that is being phosphorylated. Your answer would be a four-letter string composed of letters A and I only, e.g. IIIA. ( ) Phosphorylation of Cdc25 by M-Cdk ( ) Phosphorylation of condensin subunits by M-Cdk ( ) Phosphorylation of kinesin-5 by Aurora-A ( ) Phosphorylation of Ndc80 subunits by Aurora-B

Indicate whether each of the following descriptions better applies to a Wee1 protein (W) or a Cdc25 protein (C). Your answer would be a four-letter string composed of letters W and C only, e.g. WWCW. ( ) It is a protein kinase. ( ) It activates M-Cdk complexes. ( ) It is activated by M-Cdk complexes. ( ) Its loss in fission yeast results in small cell size.

Which division in meiosis is more similar to mitosis? In which division do sister chromatids separate from each other?

Which of the following is an inhibitory extracellular signal for cell proliferation?

Which of the following proteins is the product of an immediate early gene expressed following mitogenic stimulation of cell-cycle entry?

Mutant Saccharomyces cerevisiae cells that lack the gene encoding securin can divide more or less normally by mitosis, without significant chromosome segregation defects. Cells harboring a nondegradable version of securin, on the other hand, arrest in metaphase as expected, since they cannot activate separase to enter anaphase. Similarly, cells lacking Cdc20 arrest in metaphase, since they cannot activate APC/C. Finally, cells lacking both securin and Cdc20 arrest in anaphase: they manage to separate sister chromatids, but do not progress much further. These results suggest that in wild-type cells, …

Treatment of dividing cells with a low dose of the antifungal drug benomyl, which destabilizes microtubules, slows down correct spindle assembly. But at such doses, the spindle is eventually formed and the cells survive. However, mutations in some genes confer benomyl sensitivity: the mutant cells die because they fail to arrest the cell cycle in the presence of unattached kinetochores and progress through anaphase, with disastrous consequences. Which of the following would you expect to be a benomyl-sensitive mutant?

Which one better supports cell proliferation when added to fibroblast cultures: serum or plasma? This activity is due to the presence of mitogens in this fluid. What is responsible for making these mitogens?

To study chromosome movement during anaphase in mammals, you have injected fluorescently labeled tubulin subunits into cells such that microtubules can be seen under a fluorescence microscope. You then use a relatively strong laser beam to bleach the fluorescent dyes in a limited area of a metaphase cell, as indicated in the schematic diagram below, and follow the progression of mitosis under the microscope by time-lapse imaging. You then use the images to measure the change in distance between various components, and plot the results in the graph below. Answer the following question(s) based on these results. -Based on the results from the experiment, which is dominant in this cell: anaphase A or anaphase B? Write down A or B as your answer.

Three models for contractile-ring positioning in animal cells are presented in the schematic diagrams below. Answer the following question(s) according to these models. -In the early embryos of Caenorhabditis elegans, defects in the formation of astral microtubules increase myosin activity throughout the cell cortex. If the spindle is forced to one side of the cell, the cortical myosin activity is observed mostly at the opposite side of the cell. Which model (1, 2, or 3) better predicts these observations? Write down 1, 2, or 3 as your answer.

How is Cdc20-APC/C similar to Cdh1-APC/C?

Which of the following cell populations in our body has the highest mitotic index?

To study chromosome movement during anaphase in mammals, you have injected fluorescently labeled tubulin subunits into cells such that microtubules can be seen under a fluorescence microscope. You then use a relatively strong laser beam to bleach the fluorescent dyes in a limited area of a metaphase cell, as indicated in the schematic diagram below, and follow the progression of mitosis under the microscope by time-lapse imaging. You then use the images to measure the change in distance between various components, and plot the results in the graph below. Answer the following question(s) based on these results. -Based on the results from your experiment, which force is dominant in chromosome movement in this cell: polar ejection force (E), microtubule flux (F), or kinetochore microtubule plus-end depolymerization (P)? Write down E, F, or P as your answer.

Formin nucleates the growth of parallel actin bundles in the cell. Additionally, myosin motor activity is positively regulated by phosphorylation. The monomeric G protein RhoA is important in cytokinesis, because it directly or indirectly …

Consider two kinesin motor proteins at the mitotic spindle midzone: kinesin-5 is a tetrameric motor that walks toward the plus end of both microtubules to which it is attached via its motor domains; kinesin-14, on the other hand, walks toward the minus end of one microtubule while it is attached to another microtubule via its tail domain. How do these motors affect the length of the spindle?

Mammalian Cdk inhibitor proteins (CKIs) can be grouped into two families based on their structural and functional differences. The Cip/Kip family proteins (e.g. p21) have a broad binding specificity. These proteins bind preferentially to already formed cyclin-Cdk complexes and thus enhance complex formation. However, they inhibit the kinase activity of most complexes (e.g. S-Cdks), except in the case of G?-Cdk complexes where no inhibition occurs. Consequently, Cip/Kip family proteins have an overall positive effect on Cdk4/6 activity due to their help in bringing the subunits together. In contrast, the inhibitors of the INK4 family (e.g. p16) bind only to the Cdk subunit of G?-Cdks and prevent binding of both the G? cyclins and the Cip/Kip family CKIs. Based solely on these findings, would you expect p16 to activate (A) or inactivate (I) the S-Cdks in the presence of limited amounts of p21?

You have been studying the effect of loss-of-function mutations in the Cdk inhibitor protein (CKI) p21. You add the drug fucoxanthin to cell cultures harboring either wild-type or mutant versions of the p21 gene. Fucoxanthin is known to induce cell-cycle arrest in G1. After a day, you add the thymidine analog BrdU to the culture media, collect the cells after an hour, treat them with anti-BrdU antibody and the fluorescent DNA stain DAPI, and finally subject them to fluorescence-activated cell sorting (FACS). The FACS data can be viewed as a two-dimensional dot plot composed of thousands of dots, in which each cell is represented by one dot at coordinates that correspond to the intensities of the DAPI fluorescence signal (X axis) and BrdU fluorescence signal (Y axis) for that cell. Answer the following question(s) according to the simplified dot plot below, generated from your experiment. -Indicate which boxed region (1, 2, or 3) in the FACS plots corresponds better to each of the following phases of the cell cycle. Your answer would be a three-digit number composed of numbers 1 to 3, with each number used once, e.g. 312. ( ) G₁ phase ( ) S phase ( ) G₂ and M phases