Functional characterization of centriolar proteins CEP120 and SPICE1
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Animal cells have a cytoskeleton that gives them their shape and structure, just like the skeleton does with a whole organism. The cytoskeleton consists of three components: actin filaments, intermediate filaments and microtubules. All three components have a slightly different function, but we will focus on the microtubules in this study. As the name says, microtubules are small, hollow tubes. They are formed by the protein tubulin and stretch throughout the cell to form a network. Besides their structural function, microtubules are also important in intracellular transport. The microtubule network is organized by a small structure called the centrosome. This centrosome is located near the nucleus of the cell, and from there, the microtubules extend through the whole cell. The centrosome is formed by two centrioles that are arranged perpendicularly. Centrioles are cylinders built from microtubules and centriolar proteins. The centrioles are approximately 500 nm long in human cells. Centriolar proteins are proteins that are located at the centrioles and perform their function there. The structure of the centrosome is crucial for the microtubules to be able to grow and form the network as they should. Besides the centrosome's normal function in the cell, it also has a role in mitosis. During cell division, the centrosome is duplicated and the two centrosomes form the mitotic spindles that help divide the genome evenly over the two cells. If there is a problem with genome distribution, this can lead to issues with the newly formed cells or even cancer. There are also diseases that are caused by poorly or non-functioning centrosomes, and more specifically, problems with the centriolar proteins. Because issues with the centrosome can have disastrous results, it is vital to understand their function better. A better understanding of the centrosome could help treat diseases caused by issues with the centrosome. After each cell division, the centrosome needs to be duplicated because each daughter cell only receives one centrosome consisting of two centrioles. For a new centrosome to form, two new centrioles need to grow from the two existing centrioles. This duplication happens during a process called centriole biogenesis. It is important that this process goes well since two functioning centrosomes are crucial for further cell divisions and the functioning of the daughter cells. The centrosomal proteins mentioned before are also involved during centrosome biogenesis. The centrosome cannot assemble and thus function properly without the centriolar proteins. However, there are many different centriolar proteins and not for all of them, their role in centriole biogenesis is uncovered. In this study, we look at two centrosomal proteins called CEP120 and SPICE1. Of these two proteins, the function in the centrosome is not entirely clear yet. With different experimental techniques, we will try to elucidate the role of CEP120 and SPICE1 in the centrosome. We used the fluorescent proteins GFP and mCherry to visualize the two proteins of interest. In this way, we can observe them under a microscope and analyze their behavior. We found effects and interactions of CEP120 and SPICE1 that could be important in centriole biogenesis.