Maksim Viktorovich Imakaev

Formation of Chromosomal Domains by Loop Extrusion

Topologically associating domains (TADs) are fundamental structural and functional building blocks of human interphase chromosomes, yet the mechanisms of TAD formation remain unclear. Here, we propose that loop extrusion underlies TAD formation. In this process, cis-acting loop-extruding factors, likely cohesins, form progressively larger loops but stall at TAD boundaries due to interactions with boundary proteins, including CTCF. Using polymer simulations, we show that this model produces TADs and finer-scale features of Hi-C data. Each TAD emerges from multiple loops dynamically formed through extrusion, contrary to typical illustrations of single static loops. Loop extrusion both explains diverse experimental observations-including the preferential orientation of CTCF motifs, enrichments of architectural proteins at TAD boundaries, and boundary deletion experiments-and makes specific predictions for the depletion of CTCF versus cohesin. Finally, loop extrusion has potentially far-ranging consequences for processes such as enhancer-promoter interactions, orientation-specific chromosomal looping, and compaction of mitotic chromosomes.

High-Resolution Mapping of the Spatial Organization of a Bacterial Chromosome

Caulobacter Chromosome Chromosomal DNA must be highly compacted to fit within the tiny volume of the cell, while at the same time it must maintain a conformation that allows critical cellular processes access to the genome. Le et al. (p. 731, published online 24 October) analyzed the structure of the circular chromosome in the prokaryote Caulobacter crescentus by using chromosome conformation capture and deep-sequencing. Highly self-interacting regions (chromosomal interaction domains, or CIDs) were observed—similar to the topologically associated domains previously seen in eukaryotes. Supercoiling helped to establish CIDs, and CID boundaries were defined by highly expressed genes. CIDs appeared to be established during or shortly after DNA replication, and could potentially facilitate chromosomal segregation by preventing newly replicated chromosomes from becoming entangled. A bacterial chromosome is organized into self-interacting regions delimited by highly expressed genes. Chromosomes must be highly compacted and organized within cells, but how this is achieved in vivo remains poorly understood. We report the use of chromosome conformation capture coupled with deep sequencing (Hi-C) to map the structure of bacterial chromosomes. Analysis of Hi-C data and polymer modeling indicates that the Caulobacter crescentus chromosome consists of multiple, largely independent spatial domains that are probably composed of supercoiled plectonemes arrayed into a bottle brush–like fiber. These domains are stable throughout the cell cycle and are reestablished concomitantly with DNA replication. We provide evidence that domain boundaries are established by highly expressed genes and the formation of plectoneme-free regions, whereas the histone-like protein HU and SMC (structural maintenance of chromosomes) promote short-range compaction and the colinearity of chromosomal arms, respectively. Collectively, our results reveal general principles for the organization and structure of chromosomes in vivo.

Effects of topological constraints on globular polymers

Topological constraints can affect both equilibrium and dynamic properties of polymer systems and can play a role in the organization of chromosomes. Despite many theoretical studies, the effects of topological constraints on the equilibrium state of a single compact polymer have not been systematically studied. Here we use simulations to address this longstanding problem. We find that sufficiently long unknotted polymers differ from knotted ones in the spatial and topological states of their subchains. The unknotted globule has subchains that are mostly unknotted and form asymptotically compact RG(s)∼s1/3 crumples. However, crumples display a high fractal dimension of the surface db=2.8, forming excessive contacts and interpenetrating each other. We conclude that this topologically constrained equilibrium state resembles a conjectured crumpled globule [Grosberg et al., Journal de Physique, 1988, 49, 2095], but differs from its idealized hierarchy of self-similar, isolated and compact crumples.