Spermidine-induced aggregation of nucleosome core particles: evidence for multiple liquid crystalline phases

Abstract

We investigate the effect of the addition of a trivalent cation, spermidine, to dilute solutions of nucleosome core particles (NCP). In the presence of spermidine, part of the NCP segregates from the initial homogeneous solution, forming dense aggregates. We follow this precipitation process over a wide range of spermidine and NaCl concentrations and determine the conditions of aggregation of the particles. The structure of the dense phases is analyzed by means of polarizing light microscopy and cryo-electron microscopy. We report the existence of multiple supramolecular organizations. According to the relative concentrations of spermidine, monovalent salt and NCP, the particles may aggregate into amorphous phases, stack into randomly oriented columns, or form liquid crystalline phases. Two discotic liquid crystalline phases are identified and analyzed: a columnar nematic corresponding to columns of NCP simply aligned in parallel, and a columnar hexagonal phase in which the columns order into a transversal 2D hexagonal array. We discuss the nature and origin of the interactions possibly involved in the formation and maintenance of these different types of order.