Localization and Behavior of Nuclear Lamin Proteins
thesisposted on 14.05.2020, 00:00 authored by Dallin North
Nuclear size has been observed to be irregular in cancer cells, and as such can be used to phenotypically identify cancer cells and their stages. Many cancers have unusually large nuclei, and in certain cancers, nuclear size increases as the cancer becomes more aggressive. In order to fully understand the impact nuclear size has in cancer, and determine to what extent it is a cause or effect, we must understand how nuclear size is controlled. Although significant progress has been made over the past few years, many of the mechanisms and intricacies behind nuclear size regulation remain unknown. This project aims to determine one mechanism by which nuclear lamin proteins, which partially comprise the nucleoskeleton and help provide some structure to the organelle, help to control nuclear size. Previous research has suggested nuclear lamins may hold an important role in nuclear size regulation. Overall, this project intends to create an easily observable model system in human HeLa cells. This cell line, via CRISPR technology, will be genetically modified to endogenously tag Lamin A/C with enhanced green fluorescent protein (eGFP). This will allow Lamin A/C dynamics to be measured with Fluorescence Recovery After Photobleaching (FRAP) techniques. Additionally, as high protein kinase C (PKC) β activity has been implicated in the reduction of nuclear size in HeLa and MRC-5 fibroblast cells, we plan to introduce a constitutively active version of PKC β to view the resulting behavior on dynamics. Ideally, these experiments will shed some light on how Lamin A/C dynamics relate to nuclear size. The results may be utilized with current cancer research to hopefully provide more understanding to the way cancer works or maybe even potential treatments.