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Localization and interactions of DNA damage response and cell cycle checkpoint proteins in yeast and chicken cells using fluorescente microscopy


Cancer is a major killer in the western world, second only to heart disease as a cause of mortality. Despite its impact cancer is a poorly understood disease. Fundamental research into both the causes and mechanisms of cancer are required if the outlook for cancer patients is to be improved. Cancer is caused by an accumulation of mutations, a process referred to as tumour progression.

This accumulation is required since there are multiple control mechanisms to prevent cells becoming cancerous. Many cancers involve mutations in the DNA repair machinery allowing further mutations, leading to tumour progression and cancer. It is likely that mutations in DNA repair and cell cycle checkpoint are early steps in tumour progression. Significantly, mutations of some proteins in this study cause genetic diseases predisposing humans to cancer.

The aim of this project is to study the localisation and interactions of DNA damage repair and cell cycle checkpoint proteins after induction of DNA damage. This will be achieved by tagging selected proteins with fluorescent protein motifs and studying them using fluorescence microscopy. The large amount of conservation in the relevant cellular mechanisms allows the use of model organisms such as yeast and DT40 chicken cells.

I will initially exploit the ease of genetic manipulation in the yeast Saccharomyces cerevisiae to replace genomic copies of the proteins of interest with fully functional fusion constructs of the relevant protein with either Yellow or Cyan Fluore scent Protein (YFP or CFP). Initial studies will focus on localisation of single proteins after DNA damage, followed by Co-localisation of protein pairs. Interesting interactions detected in yeast experiments will be further studied in chicken DT40 cells.

Call for proposal

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Funding Scheme

EIF - Marie Curie actions-Intra-European Fellowships


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