Bio471 Aaron Broege, PhD Literature Module #1 Peer-reviewed article Title: The Breast Cancer Susceptibility Gene BRCA1

1 Is Required for Subnuclear Assembly of Rad51 and Survival following Treatment with the DNA Cross-linking Agent Cisplatin. Primary author: Anamitra Bhattacharyya Journal: The Journal of Biological Chemistry INSTRUCTIONS: All students will read the entire article; however, each individual will be responsible for answering a set of questions specific to one figure of the paper. Everyone will answer the general questions section. In class, you will meet with the individuals from other home groups that were assigned the same figure and reach a consensus of what you gleaned from the figure. Doing this provides an opportunity for you to share with classmates points on which you still may be uncertain. Following this, you will return to your home groups and each individual will take turns explaining their figure, and the group will discuss how the paper fits together as a whole. General (everyone answer) Understanding definitions of these proteins, molecules, etc. will increase your ability to understand the paper. 1. Describe the molecular function of BRCA1 and BRCA2.

2. Describe the molecular function of Rad51.

3. Describe the effects of cisplatin treatment and x-ray exposure on DNA.

4. What does the term knockout mouse mean?

5. State the authors hypothesis. (Hint: The hypothesis will typically be clearly stated at the end of the introduction)

Bio471 Aaron Broege, PhD Figure 1 (Individual A is responsible for this section) 1. What question are the authors addressing in this figure?

2. Cells in figure 1 were stained with 4,6-diamidino-2-phenylindole, or DAPI (you will commonly see this referred to as just DAPI). What purpose does DAPI staining serve?

3. In addition to DAPI, these cells have been treated with another reagent for visualizing a specific protein. What else is visualized in this figure and why are they looking at that particular protein?

4. What do the foci in this figure represent?

5. Why are the authors comparing brca+/+ and brca-/- cells?

6. What conclusion do the authors draw from this figure? Do you agree that their data support their conclusion?

Figure 2 (Individual B is responsible for this section) 1. What question(s) are the authors addressing in this figure?

Bio471 Aaron Broege, PhD 2. Describe the approach they take to answering this question.

3. What cell types were used in this figure? Why were the two different cell types used?

4. Compare and contrast panel A and panel B. What are they testing in each? What information does each panel tell them?

5. What two conclusions do the authors draw from this figure? Do you agree that the data support these conclusions?

Figure 3, Table 1, and Figure 4 (Individual C is responsible for this section) Figure 3/Table 1 (Table 1 is quantification of the data in Figure 3) 1. What question are the authors attempting to answer in figure 3?

2. What technique do the authors use to answer this question?

Bio471 Aaron Broege, PhD 3. What do the authors conclude in figure 3/table 1? Do you agree that the data supports their conclusion?

Figure 4 4. What question are the authors asking in Figure 4? What is their reason for testing this question?

5. What technique are they using to address their question? What is this technique used to test?

6. What conclusion do the authors draw from this figure? Do you agree that the data supports that conclusion?