Small T Antigen Effect on Mitotic Proteins B-Myb and FOXM1
Mentor: James A. DeCaprio, MD (Dana Farber Cancer Institute)
Merkel Cell Carcinoma (MCC) is a rare and aggressive skin cancer with a high rate of metastasis and death. Though MCC itself is quite rare, Merkel cell polyomavirus (MCPyV) is found in a large percentage of the human population. Furthermore, it has been found that 80% of MCCs are positive for MCPyV. Polyomaviruses are characterized by their non-enveloped double stranded DNA that code for both large and small T proteins. The small T antigen is commonly associated with the upregulation of late cell cycle genes (G2/M phase). Similarly, N-MYC and C-MYC are genes which are believed to regulate FOXM1 and B-Myb respectively. Both are part of a protein complex that pushes cells into mitosis. My research will also involve the introduction of the aniPOND technique to the lab. AniPOND is essentially the inverse of ChIP-Sequencing. By pulling down the DNA and looking at the proteins attached using aniPOND, I will investigate small T’s effect on FOXM1 and B-Myb.
HeLa cells were synchronized using a double thymidine block, causing them to progress through the cell cycle at a similar rate. The success of this synchronization is measured using flow cytometry. By evaluating the size and consistency of the cells, the machine is able to help us identify which stage of the cell cycle the sample was in. Pictured is the histogram of the 4 hour time point. At this time a majority of the cells were in S phase, proving the synchronization to be a success. Source: