You have isolated a temperature-sensitive cell-cycle mutant. The mutated gene encodes the activating subunit of the APC. The mutant protein is defective at high temperatures. You sort several thousand cells and recover the WT-looking profile below: Cells (in thousands) O G1 COM O m OS N Next you grow the mutant at the restrictive high temperature and sort an equal number of cells. First, what phase of the cell cycle do you expect the cells to arrest in? T 50 100 150 200 Propidium iodide fluorescence 250 #tv A @ X O
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- You have isolated a temperature-sensitive cell-cycle mutant. The mutated gene encodes the activating subunit of the APC. The mutant protein is defective at high temperatures. You sort several thousand cells and recover the WT-looking profile below: Cells (in thousands) O G1 OM O G2 OS E 5 Next you grow the mutant at the restrictive high temperature and sort an equal number of cells. First, what phase of the cell cycle do you expect the cells to arrest in? 3 2 50 100 150 200⁰ 250 Propidium iodide fluorescence ♫ #tv A X MacBook Airin term of signal transduction how cell cycle/division is controlled?? ((while you are answerig plz write the refrence.))Cell division cycle mutations render the mutants unable to continue the cell cycle. This phenotype creates a paradox where mutant cells must also be grown in the lab to further identify the gene and study the role of the protein. How do you think this problem can be solved?
- One approach to studying the regulation of cell cycle progression (particularly in an era when genetic and molecular biology manipulations were less readily accomplished in mammalian cells) was to use treatments that induced cells to fuse and then monitor the behavior of the two nuclei in the resulting cell. The figure below depicts data from one such study. The investigators did preliminary work to produce populations of cells that were synchronized in various stages of the cell cycle (G1, S, or G2 in the examples shown below). They then fused the cells in different combinations and monitored subsequent events in each of the nuclei. For purposes of this question, we will pay particular attention to what occurred in the nucleus that came from the cell in G1. In one experiment (I), cells in the G1 and S phases were fused. That event caused the nucleus from the G1 cell to very quickly enter the S phase (sooner than it would otherwise have done so). In contrast, in a second experiment…In order to investigate the cell cycle in HEK293 cells in culture, I ran a flow cytometry experiment. I labeled their nuclei with DAPI to analyze DNA content. In these flow cytometry plots below, label the two main phases of the cell cycle that you can see. Briefly, explain your answer.One approach to studying the regulation of cell cycle progression (particularly in an era when genetic and molecular biology manipulations were less readily accomplished in mammalian cells) was to use treatments that induced cells to fuse and then monitor the behavior of the two nuclei in the resulting cell. The figure below depicts data from one such study. The investigators did preliminary work to produce populations of cells that were synchronized in various stages of the cell cycle (G1, S, or G2 in the examples shown below). They then fused the cells in different combinations and monitored subsequent events in each of the nuclei. For purposes of this question, we will pay particular attention to what occurred in the nucleus that came from the cell in G1. In one experiment (I), cells in the G1 and S phases were fused. That event caused the nucleus from the G1 cell to very quickly enter the S phase (sooner than it would otherwise have done so). In contrast, in a second experiment…
- Biologists have long been interested in the effects of radiation on cells. In one experiment, researchers examined the effect of radium on mitosis of chick embryo cells growing in culture. A population of experimental cells was examined under the microscope for the number of cells in telophase (as a measure of mitosis occurring) before, during, and after exposure to radium. The results are shown in the Figure. What is the effect of radium exposure on mitosis? Source: R. G. Canti and M. Donaldson. 1926. The effect of radium on mitosis in vitro. Proceedings of the Royal Society of London, Series B, Containing Papers of a Biological Character 100:413419.Figure 28.11 depicts the eukaryotic cell cycle. Many cell types “exit� the cell cycle and don’t divide for prolonged periods, a state termedG0; some, for example neurons, never divide again. a. In what stage of the cell cycle do you suppose a cell might be when it exits the cell cycle and enters G0? b. The cell cycle is controlled by checkpoints, cyclins, and CDKs. Describe how biochemical events involving cyclins and CDKs might control passage of a dividing cell through the cell cycle.Passage of cells from G1 through R to S depends on the interaction of various signal molecules, proteins and enzymes which regulate the cell cycle. Imagine I have just induced a mutation in a cell line which prevents the breakdown of the cyclin molecule we discussed. Which of the following events do you predict will happen? circle all that apply Select one or more: a. Uncontrolled cell replication may result b. Retinoblastoma protein will be continuously active c. The cells will be stuck in G1 phase d. The cells will begin to replicate their DNA e. Cyclin dependent kinase (Cdk) will be continuously active
- Regulation of the cell cycle is very complex and involves multiple proteins. In yeast, a complex of cdc2 and a mitotic cyclin is responsible for moving the cell past the G2/M checkpoint. The activity of the cyclin-dependent kinase cdc2 is inhibited when it is phosphorylated by the kinase, Wee-1. What would you predict would be the phenotype of a Wee-1 mutant yeast? What other genes could be altered in a Wee-1 deficient mutant strain that would make the cells act normally?2.) One experimental tool used in the biological research and in clinical settings is called Fluorescent Activated Cell Sorting (FACS) or cell cytometry. For example, a cell membrane permeable (or nonpolar) chemical called Hoescht 33342 dye will specifically label DNA. Interpret the graph (Figure 17-4) that illustrates normal cells that have been labeled with Hoescht 33342 dye, and assign which phase(s) of the cell cycle (G1, S, G2, or M) corresponds to the fluorescence intensity. Justify your answers. 1 relative fluorescence per cell Figure 17-4 Analysis of Hoechst 33342 fluorescence in a population of cells sorted in a flow cytometer (Problem 17-15). number of cellsExplain why we can say that M-phase of the cell-cycle is triggered by a positive feedback loop. a) What would the consequences be if cohesins were working normally but condensins were not? and b) what stage of the cell cycle would this cause problems in? Why is it important for the centrosome to duplicate during G1-G2 (interphase) before M phase? The kinetochores serve as a link between the sister chromatids and the microtubules attached to the mitotic spindle. a) How are microtubules still able to exhibit dynamic instability after they are bound to the sister chromatids and b) why is this important to mitosis? As the name suggests, the Anaphase-promoting-complex (APC), promotes the 4th phase of mitosis by separating the sister chromatids so they can travel to separate poles of the cell, and prevents them from being re-zipped together. Describe how APC does these two things (Hint: one involves M-cyclin and the other involves…