The Centromeric Nucleosome in Yeast: Interaction With the Inner Kinetochore and Assembly by the Chaperone Yta7/ ATAD2
The faithful transmission of genetic information from mother to daughter cells during mitosis requires that the sister chromatids of each chromosome are properly segregated to the daughter cells. For this, the kinetochore attaches on one side to the centromeric chromatin and on the other side to the microtubule and thus provides a physical link that allows the chromatids to be pulled towards opposite poles of the cell during chromosome segregation. In the last grant period, we have characterized the attachment of the inner kinetochore to the centromeric nucleosome in the yeast Saccharomyces cerevisiae.
The current project proposes to build on this work and to obtain molecular insights into how Okp1 CENP Q/ Ame1 CENP-U interacts with the Cse4 N-terminus. Specifically, we will use proximity information obtained using biorthogonal amino acids and cross-linking/ mass spectrometry as well as structural analysis to determine the molecular details of the interaction between the centromeric nucleosome of yeast and Okp1/ Ame1 of the inner kinetochore. This is important in order to understand how physical attachment of the kinetochore to chromatin is achieved. Furthermore, we will elucidate in mechanistic detail how Yta7 ATAD2 regulates centromeric chromatin. Altogether, this project will provide mechanistic insights into regulation at the centromere that are important to understand how defects in this process contribute to chromosome missegregation.
The faithful transmission of genetic information from mother to daughter cells during mitosis requires that the sister chromatids of each chromosome are properly segregated to the daughter cells. For this, the kinetochore attaches on one side to the centromeric chromatin and on the other side to the microtubule and thus provides a physical link that allows the chromatids to be pulled towards opposite poles of the cell during chromosome segregation. In the last grant period, we have characterized the attachment of the inner kinetochore to the centromeric nucleosome in the yeast Saccharomyces cerevisiae.
The current project proposes to build on this work and to obtain molecular insights into how Okp1 CENP Q/ Ame1 CENP-U interacts with the Cse4 N-terminus. Specifically, we will use proximity information obtained using biorthogonal amino acids and cross-linking/ mass spectrometry as well as structural analysis to determine the molecular details of the interaction between the centromeric nucleosome of yeast and Okp1/ Ame1 of the inner kinetochore. This is important in order to understand how physical attachment of the kinetochore to chromatin is achieved. Furthermore, we will elucidate in mechanistic detail how Yta7 ATAD2 regulates centromeric chromatin. Altogether, this project will provide mechanistic insights into regulation at the centromere that are important to understand how defects in this process contribute to chromosome missegregation.
Duration of project
Start date: 05/2022
End date: 04/2025
Research Areas
Cell Biology, General Genetics and Functional Genome Biology
Research Areas
Genetik
