Topological domain structure of the Escherichia coli chromosome.
The circular chromosome of Escherichia coli is organized into independently supercoiled loops, or topological domains. We investigated the organization and size of these domains in vivo and in vitro. Using the expression of >300 supercoiling-sensitive genes to gauge local chromosomal supercoiling, we quantitatively measured the spread of relaxation from double-strand breaks generated in vivo and thereby calculated the distance to the nearest domain boundary. In a complementary approach, we gently isolated chromosomes and examined the lengths of individual supercoiled loops by electron microscopy. The results from these two very different methods agree remarkably well. By comparing our results to Monte Carlo simulations of domain organization models, we conclude that domain barriers are not placed stably at fixed sites on the chromosome but instead are effectively randomly distributed. We find that domains are much smaller than previously reported, approximately 10 kb on average. We discuss the implications of these findings and present models for how domain barriers may be generated and displaced during the cell cycle in a stochastic fashion.
Postow
Lisa
L
Department of Molecular and Cell Biology and Department of Mathematics, University of California at Berkeley, Berkeley, California 94720, USA.
Hardy
Christine D
CD
Arsuaga
Javier
J
Cozzarelli
Nicholas R
NR
D015139:Blotting, Southern; D002876:Chromosomes, Bacterial / Q000235:genetics* / Q000648:ultrastructure*; D003198:Computer Simulation; D004278:DNA, Superhelical / Q000235:genetics / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D008854:Microscopy, Electron; D008957:Models, Genetic; D009010:Monte Carlo Method; D009690:Nucleic Acid Conformation; D020411:Oligonucleotide Array Sequence Analysis; D015183:Restriction Mapping
D004278:DNA, Superhelical; D015246:Deoxyribonuclease EcoRI