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Investigating the relation between INI-1 and mononucleosome movement

Maguire, Brianna

Atypical teratoid/rhabdoid tumors (AT/RT) is a type of cancer that is typically seen in pediatrics. AT/RT is known to contain a mutated version of INI-1. INI-1 is a core protein within the SWI/SNF chromatin remodeling complex whose function is to move a mononucleosome up and down the strand of DNA (Allen). INI-1 has the ability to bind both to the mononucleosome DNA and to free DNA, which has already been proven by experiments facilitated within our lab. Once bound, the SWI/SNF chromatin remodeling complex then has the ability to move the mononucleosome along a strand of DNA which is important in cells, as it allows DNA to be properly read. I hypothesize that due to its role within the SWI/SNF chromatin remodeling complex and the mutations that come from INI-1 to create AT/RT cancer, INI-1 will play a vital role in proper mononucleosome movement via the SWI/SNF chromatin remodeling complex. This was tested through plasmid preparation, polymerase chain reaction, nucleosome assembly dialysis in order to assemble mononucleosomes to test the hypothesis, and then was continued through enzyme digestion and SWI/SNF remodeling to further understand mononucleosome movement. The results are expressed through various agarose and native PAGE gels to allow visualization of the length and location of the mononucleosome. To proceed further, the SWI/SNF remodeling will need to be repeated on the same in vitro mononucleosomes by using SWI/SNF without INI-1 to understand the differences and how it could possibly lead to further cancerous mutations.