Centromeres are vital structures on eukaryotic chromosomes that are essential for proper chromosome segregation during cell division. Centromeric DNA is bound by a centromere-specific variant of histone H3 known as CENP-A that replaces canonical H3 in centromeric chromatin, thus specifying centromere identity. The regulation of CENP-A chromatin assembly, however, remains poorly understood. Here, I propose experiments directed towards understanding how CENP-A chromatin is established and faithfully propagated in the fission yeast Schizosaccharomyces pombe.In Aim 1, I will adopt biochemical and genetic approaches to identify novel factors that contribute to CENP-A chromatin integrity. I expect to discover proteins that could influence CENP-A chromatin at several levels, including CENP-A assembly factors & chaperones, chromatin remodelling and histone modifying enzymes. In Aim 2, I propose to systematically characterise the novel factors identified from Aim 1, in order to assess their exact functions at centromeres. I will also perform experiments that will allow me to classify them into two distinct groups: CENP-A incorporation specific and CENP-A propagation specific factors. In Aim 3, I will devise a specialised approach to investigate the features of CENP-A that confer it the unique ability to serve as an epigenetic mark that maintains centromere position and identity.Through the state-of-the-art approaches I propose in this application, I expect to uncover the complex mechanisms that define centromeric chromatin integrity. This research will elevate Europe's reputation in epigenetics and chromatin research, and thus enhance European excellence in the life sciences.