Heterogeneous non-canonical nucleosomes predominate in yeast cells in situ

Abstract

Nuclear processes depend on the organization of chromatin, whose basic units are cylinder-shaped complexes called nucleosomes. A subset of mammalian nucleosomes in situ (inside cells) resembles the canonical structure determined in vitro 25 years ago. Nucleosome structure in situ is otherwise poorly understood. Using cryo-ET and 3-D classification analysis of yeast cells, here we find that canonical nucleosomes account for less than 10% of total nucleosomes expected in situ. In a strain in which H2A-GFP is the sole source of histone H2A, class averages that resemble canonical nucleosomes both with and without an extra density are found ex vivo, but not in situ. These data suggest that the yeast intranuclear environment favors multiple non-canonical nucleosome conformations. Using the structural observations here and the results of previous genomics and biochemical studies, we propose a model in which the average yeast nucleosome’s DNA is partially detached in situ.

Nuclear pores as conduits for fluid flow during osmotic stress

“…2d,e). While some repetitive features of this arrangement approximately matched the dimension of nucleosomes (10-12 nm x 4-6 nm) 58 , we were not able to obtain a conclusive subtomogram average of this structure from our dataset. Left panels show two tomographic slices with nuclear fibers (White arrows point to fibers).…”