Three-dimensional organization of chromatin across the plant cell cycle

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As the macromolecule complex that packages meters of genetic information in the micrometer-volume of a cell, chromatin have been under intense investigation for more than a century. Definite knowledge about the internal organization of human chromosomes and its relation to genome function has emerged recently, but much remains to be learned about the variation of chromatin folding along the cell cycle and between species. Methodological advances in cytological imaging and genomic assays have made it possible to interrogate chromatin contact probabilities and three-dimensional distances between genomic loci with high density and resolution. Our preliminary data on chromatin contact frequency at megabase resolution in barley mitotic chromosomes suggest fundamental differences in metaphase chromosome organization between Triticeae species and mammals. We propose to study chromatin folding throughout the cell cycle of barley using chromosome conformation capture sequencing, super-resolution microscopy, fluorescence in situ hybridization and computational polymer modeling. We will test whether chromatin adopts a regular solenoidal conformation in mitotic barley chromosomes. The proposed research will derive a experimentally validated structural model of chromosome organization in barley and elucidate conformational differences between large plant and smaller mammalian chromosomes.